Rgs13 Constrains Early B Cell Responses and Limits Germinal Center Sizes
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{"title"=>"Rgs13 Constrains Early B Cell Responses and Limits Germinal Center Sizes", "type"=>"journal", "authors"=>[{"first_name"=>"Il Young", "last_name"=>"Hwang", "scopus_author_id"=>"9637449400"}, {"first_name"=>"Kyung Sun", "last_name"=>"Hwang", "scopus_author_id"=>"57197884868"}, {"first_name"=>"Chung", "last_name"=>"Park", "scopus_author_id"=>"35269750100"}, {"first_name"=>"Kathleen A.", "last_name"=>"Harrison", "scopus_author_id"=>"57196683234"}, {"first_name"=>"John H.", "last_name"=>"Kehrl", "scopus_author_id"=>"7005076525"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84875285137", "sgr"=>"84875285137", "pui"=>"368580372", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"23533672", "doi"=>"10.1371/journal.pone.0060139"}, "id"=>"b14fbf14-69c5-3636-930c-255bd08a54e6", "abstract"=>"Germinal centers (GCs) are microanatomic structures that develop in secondary lymphoid organs in response to antigenic stimulation. Within GCs B cells clonally expand and their immunoglobulin genes undergo class switch recombination and somatic hypermutation. Transcriptional profiling has identified a number of genes that are prominently expressed in GC B cells. Among them is Rgs13, which encodes an RGS protein with a dual function. Its canonical function is to accelerate the intrinsic GTPase activity of heterotrimeric G-protein α subunits at the plasma membrane, thereby limiting heterotrimeric G-protein signaling. A unique, non-canonical function of RGS13 occurs following translocation to the nucleus, where it represses CREB transcriptional activity. The functional role of RGS13 in GC B cells is unknown. To create a surrogate marker for Rgs13 expression and a loss of function mutation, we inserted a GFP coding region into the Rgs13 genomic locus. Following immunization GFP expression rapidly increased in activated B cells, persisted in GC B cells, but declined in newly generated memory B and plasma cells. Intravital microscopy of the inguinal lymph node (LN) of immunized mice revealed the rapid appearance of GFP(+) cells at LN interfollicular regions and along the T/B cell borders, and eventually within GCs. Analysis of WT, knock-in, and mixed chimeric mice indicated that RGS13 constrains extra-follicular plasma cell generation, GC size, and GC B cell numbers. Analysis of select cell cycle and GC specific genes disclosed an aberrant gene expression profile in the Rgs13 deficient GC B cells. These results indicate that RGS13, likely acting at cell membranes and in nuclei, helps coordinate key decision points during the expansion and differentiation of naive B cells.", "link"=>"http://www.mendeley.com/research/rgs13-constrains-early-b-cell-responses-limits-germinal-center-sizes", "reader_count"=>18, "reader_count_by_academic_status"=>{"Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>2, "Student > Master"=>6, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>2, "Student > Master"=>6, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>14, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>1, "Italy"=>1, "Germany"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/995754"], "description"=>"<p>A. Intravital TP-LSM of the inguinal LN of KI mice immunized with sRBCs 1, 2, or 4 days previously. The day prior to imaging labeled B cells or labeled B and T cells from non-immunized WT mice were transferred to outline LN follicle and T cell zone, respectively. Shown are X, Y; X, Z (below); and Y, Z (right) projections from an image stack collected on the indicated day. Clusters of GFP positive cells shown with white arrow and GFP positive cells in the follicle delineated by yellow arrows. B. Intravital TP-LSM of the inguinal LN of a KI mouse immunized with sRBC 7 days previously. The day prior to imaging splenic B cells from a non-immunized WT mouse were adoptively transferred to outline the LN follicle. The left image shows GFP expression, the transferred WT B cells (red), and collagen (blue). The middle two images show the region subjected to analysis and tracks of the WT B cells (red) and the endogenous GFP expressing cells present in the KI mice. C. Track analysis of GFP<sup>+</sup> KI B cells in the GC region versus WT B cells within the follicle. Statistical significance of straightness and displacement was calculated by Mann Whitney test. (*; p<0.05, **; p<0.01) D. Electronically zoomed time lapse images from GFP<sup>+</sup> KI B cell in the GC from part B intravital TP-LSM imaging.</p>", "links"=>[], "tags"=>["induction", "gfp"], "article_id"=>658509, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g004", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Rapid_induction_of_GFP_expression_in_vivo_following_immunization_/658509", "title"=>"Rapid induction of GFP expression <i>in vivo</i> following immunization.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:18:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/995747"], "description"=>"<p>A. RT-PCR analysis of RNA extracted from sorted B cell population obtained from mice 8 days after sRBC immunization. B. Quantitative RT-PCR analysis of sorted B cell populations from immunized and non-immunized mice. C. Microarray data analysis of <i>Rgs13</i> expression in sorted cell populations. Data extracted from the Immunological Genome Project database (<a href=\"http://www.immgen.org/databrowser/index.html\" target=\"_blank\">http://www.immgen.org/databrowser/index.html</a>) D. Brightfield microscopy of a sectioned Peyer's Patch prepared from WT and KI mice Immunostained for RGS13, CD4, and IgD. Scale bar, 50 µm, left panel; and 3X zoom, right panel. An insert in the right panel is from the GC region of a similarly immunostained KI mouse. E. Immunoblot analysis of RGS13 expression in WT and KI immunized splenocytes prepared from day 10 immunized WT and KI mice. Actin levels were used as a loading control. F. Confocal microscopy of Peyer's Patch cells prepared from WT and KI mice immunostained for RGS13 and B220. Individual and merged images are shown. Scale bar, 7 µm. G. Confocal microscopy of GFP positive B cell prepared from D7 immunized KI mice.</p>", "links"=>[], "tags"=>["immunology"], "article_id"=>658502, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g001", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_RGS_protein_gene_expression_in_various_B_cell_populations_/658502", "title"=>"RGS protein gene expression in various B cell populations.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:15:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/995762"], "description"=>"<p>A. ELISA assay results from analysis of sera collected at the indicated days from WT and KI mice immunized subcutaneously with TNP-KLH in complete Freund's adjuvant and boosted at day 28. IgM, IgG1, IgG2b, IgG2c and IgG3 specific antibodies at each time point were assayed with plates coated with TNP<sub>34</sub>BSA or TNP<sub>3</sub>BSA (left panels). The ratios between the TNP<sub>3</sub> and TNP<sub>34</sub> responses are shown (right panels) for the WT and KI mice. Results are from 4 WT versus 4 KI mice. Similar results were obtained from 2 additional experiments. B. ELISPOT assay results from analysis of spleen cells from 2 WT and 2 KI mice immunized 5 days previously with TNP-KLH. Similar results in 2 other experiments. Data is mean ± SEM and statistics from unpaired t tests. C. ELISPOT assay results from analysis of bone marrow, blood, and spleen cells at the indicated time points following immunization with TNP-KLH. The numbers of TNP specific ELISPOTs are shown from 1 experiment comparing 2 WT versus 2 KI mice. Similar results from one other experiment. Data is mean ± SEM and statistics from unpaired t tests. D. ELISPOT assay results from analysis of spleen cells from 2 WT and 2 KI mice immunized 6 days previously with sRBCs. Similar results in 1 other experiment. Data is mean ± SEM and statistics from unpaired t tests. E. Representative brightfield microscopy images of WT and KI spleen sections immunostained for IgG (blue) and B220 (brown). F. Flow cytometric quantification of the number of CD138<sup>+</sup>B220<sup>+</sup> and CD138<sup>+</sup>B220<sup>−</sup> B cells at D11 post-immunization with sRBCs in the spleens, axillary and inguinal LNs, mesenteric LNs, and Peyer's Patches of 4 mice reconstituted with a 1∶1 mix of WT and KI bone marrow 8 weeks after reconstitution. Data is mean ± SEM and statistics from unpaired t tests.</p>", "links"=>[], "tags"=>["antibody", "ki"], "article_id"=>658513, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g007", "stats"=>{"downloads"=>3, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Enhanced_early_antibody_response_in_the_Rgs13_GFP_KI_mice_/658513", "title"=>"Enhanced early antibody response in the <i>Rgs13</i>GFP KI mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:21:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/995756"], "description"=>"<p>A. Brightfield microscopy of representative spleen sections from day 9 and 30 sRBC immunized WT and KI mice using antibodies against CD35 and Ki67, IgD and CD3, or IgM and IgD. In some sections GCs are indicated with arrowheads. B. Quantification of the number of GCs per spleen section from WT and KI mice 8–10 or 30 days post-immunization with sRBCs. Eight WT and 8 KI mice (8–10 day) and 4 WT and 4 KI mice (day 30) immunostained for Ki67 and CD35 were used. Data is mean ± SEM. Statistics, unpaired t test. C. Quantification of CD35 and Ki67 immunostaining of individual GCs from WT and KI spleen sections prepared from 8–10 day post immunized animals. Data is mean ± SEM of the areas from CD35 and Ki67 immunostaining of 50 WT and KI GCs (unpaired t test). D. Flow cytometric analysis of B220<sup>+</sup>CD38<sup>−</sup>GL7<sup>+</sup>CD95<sup>+</sup> cells in Peyer's patches from WT and KI mice prior to and post sRBC immunization. Data is % of B220 gate and is the mean ± SEM of 8 v. 8, 11 v. 11, 8 v. 8, and 4 v. 4 WT and KI mice at 0, 3–5, 10–11, and 30 days post immunization, respectively. Results compared by unpaired t test. E. Brightfield microscopy of representative mesenteric LNs from WT and KI mice using antibodies against CD35 and Ki67. F. Flow cytometric analysis of B220<sup>+</sup>CD38<sup>−</sup>GL7<sup>+</sup>CD95<sup>+</sup> cells in mesenteric and peripheral LNs from WT and KI mice prior to and post sRBC immunization. Data is % of B220 gate and is the mean ± SEM of 4 WT v. 4 KI mice at each time point. Results compared by unpaired t test. G. Flow cytometric enumeration of B220<sup>+</sup>CD38<sup>−</sup>GL7<sup>+</sup>CD95<sup>+</sup> cells in peripheral LNs (pLN), mesenteric LNs (mLN), Peyer's patches (PP) and the spleen from chimeric mice (CD45.1 versus CD45.2) 9 days post sRBC immunization. Data is mean ± SEM of cells recovered from 4 chimeric mice. Results are compared by unpaired t test.</p>", "links"=>[], "tags"=>["gcs", "ki"], "article_id"=>658511, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g005", "stats"=>{"downloads"=>2, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Large_GCs_in_Rgs13_GFP_KI_mice_/658511", "title"=>"Large GCs in <i>Rgs13</i>GFP KI mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:20:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/995759"], "description"=>"<p>A, Chemotaxis assays of WT and KI spleen cells from day 10 sRBC immunized mice immunostained with B220, GL7, and CD95 using various concentrations of CXCL12, CXCL13, or CCL19. In the last panel the KI GC B cells were further fractionated on the basis of GFP expression. Results are from the analysis of 4 WT and 4 KI mice with each assay preformed in triplicate. Data is mean ± SEM and statistics from unpaired t tests. B. Chemotaxis assays of WT and KI LN cells from D10 sRBC immunized mice immunostained with B220, CD4, and IgD using various concentrations to CCL19 or CXCL13. Results are from the analysis of 2 WT and 2 KI mice with each assay preformed in triplicate. Data is mean ± SEM. Experiment repeated twice with similar results. C. Chemotaxis assay of WT and KI Peyer's Patch cells immunostained with B220, GL7, and CD95 to indicated concentration of CXCL12, CXL13, or CCL19. The KI GC B cells were fractionated based on GFP expression. Results are from the analysis of 1 WT and 1 KI mice with each assay preformed in quadruplicate. Data is mean ± SEM and statistics from unpaired t test. Similar results in 3 other experiments. D. Chemotaxis assays of WT (CD45.2) and KI (CD45.1) spleen cells from day 10 sRBC immunized chimeric mice immunostained with B220, CD38, GL7, and CD95 using indicated concentrations of CXCL12, CXCL13, or CCL19. Results are from the analysis of 4 chimeric mice with each assay performed in triplicate. Data is mean ± SEM and statistics from unpaired t tests.</p>", "links"=>[], "tags"=>["cells", "wt", "ki", "mice", "responses"], "article_id"=>658512, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g006", "stats"=>{"downloads"=>0, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GC_B_cells_from_WT_and_Rgs13_GFP_KI_mice_exhibit_similar_responses_to_chemokines_/658512", "title"=>"GC B cells from WT and <i>Rgs13</i>GFP KI mice exhibit similar responses to chemokines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:20:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/995752"], "description"=>"<p>A. Flow cytometric quantification of absolute number of B220<sup>+</sup>CD38<sup>−</sup> cells in the spleens of WT versus KI mice at various days post sRBC immunization. B. Flow cytometric quantification of absolute number of B220<sup>+</sup>CD38<sup>+</sup>GL7<sup>+</sup> cells in the spleens of WT versus KI mice at various days post sRBC immunization. C. Flow cytometric quantification of absolute number of GC B cells in the spleens of WT versus KI mice at various days post sRBC immunization. D. Flow cytometric quantification of absolute number of early plasma cells in the spleens of WT versus KI mice post immunization. E. Flow cytometric quantification of absolute number of mature plasma cells in the spleens of WT versus KI mice following immunization. F. Flow cytometric quantification of absolute number of B220<sup>+</sup>CD38<sup>+</sup>IgG1<sup>+</sup> cells in the spleens of WT versus KI mice post immunization. Results are based on the analysis of 6 WT versus 6 KI mice at each time point. The absolute cell number of B-cell subsets & plasma cells (CD138<sup>+</sup>) were calculated from the flow cytometric analysis and presented as the mean ± SEM of six per group. Statistics are from unpaired t tests.</p>", "links"=>[], "tags"=>["numbers", "cells", "immunization", "ki"], "article_id"=>658507, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g003", "stats"=>{"downloads"=>2, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Increased_numbers_of_GC_plasma_and_memory_B_cells_following_immunization_of_the_Rgs13_GFP_KI_mice_/658507", "title"=>"Increased numbers of GC, plasma, and memory B cells following immunization of the <i>Rgs13</i>GFP KI mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:17:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/995751"], "description"=>"<p>A. Representative flow cytometry plots of spleen cells prepared from KI mice prior to and at days 2, 5, and 11 post sRBC immunization using antibodies specific for B220, CD38, CD95 (FAS), and GL7 along with GFP. Cells were gated as indicated above each plot and GFP expression is shown on the x-axis. B. Representative flow cytometry plots of spleens cells prepared from KI mice at day 11 post immunization using the same antibodies as part A along with antibodies specific for CXCR4 and CD86. Dark zone cells are GC B cells that are CXCR4<sup>hi</sup>CD86<sup>low</sup> while light zone cells are CXCR4<sup>low</sup>CD86<sup>hi</sup>. C. Representative flow cytometry plots of spleens cells prepared from KI mice 5, 11, or 30 days post sRBC immunization using antibodies specific for B220, CD38, and IgG1 along with GFP. Cells gated as indicated. D. Representative flow cytometry plots of spleen cells prepared from KI mice at 5 or 11 days post immunization using antibodies specific for B220 and CD138 along with GFP. Cells gates are as indicated. All experiments preformed a minimum of 3 times with 2–4 mice and presented as the mean ± SEM of 2–4 mice per group.</p>", "links"=>[], "tags"=>["cytometric", "gfp", "immunized", "ki"], "article_id"=>658506, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g002", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Flow_cytometric_analysis_of_GFP_expression_in_immunized_Rgs13_GFP_KI_mice_/658506", "title"=>"Flow cytometric analysis of GFP expression in immunized <i>Rgs13</i>GFP KI mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:17:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/995771", "https://ndownloader.figshare.com/files/995773", "https://ndownloader.figshare.com/files/995774", "https://ndownloader.figshare.com/files/995776", "https://ndownloader.figshare.com/files/995778", "https://ndownloader.figshare.com/files/995779"], "description"=>"<div><p>Germinal centers (GCs) are microanatomic structures that develop in secondary lymphoid organs in response to antigenic stimulation. Within GCs B cells clonally expand and their immunoglobulin genes undergo class switch recombination and somatic hypermutation. Transcriptional profiling has identified a number of genes that are prominently expressed in GC B cells. Among them is R<i>gs13</i>, which encodes an RGS protein with a dual function. Its canonical function is to accelerate the intrinsic GTPase activity of heterotrimeric G-protein α subunits at the plasma membrane, thereby limiting heterotrimeric G-protein signaling. A unique, non-canonical function of RGS13 occurs following translocation to the nucleus, where it represses CREB transcriptional activity. The functional role of RGS13 in GC B cells is unknown. To create a surrogate marker for <i>Rgs13</i> expression and a loss of function mutation, we inserted a GFP coding region into the <i>Rgs13</i> genomic locus. Following immunization GFP expression rapidly increased in activated B cells, persisted in GC B cells, but declined in newly generated memory B and plasma cells. Intravital microscopy of the inguinal lymph node (LN) of immunized mice revealed the rapid appearance of GFP<sup>+</sup> cells at LN interfollicular regions and along the T/B cell borders, and eventually within GCs. Analysis of WT, knock-in, and mixed chimeric mice indicated that RGS13 constrains extra-follicular plasma cell generation, GC size, and GC B cell numbers. Analysis of select cell cycle and GC specific genes disclosed an aberrant gene expression profile in the <i>Rgs13</i> deficient GC B cells. These results indicate that RGS13, likely acting at cell membranes and in nuclei, helps coordinate key decision points during the expansion and differentiation of naive B cells.</p> </div>", "links"=>[], "tags"=>["rgs13", "constrains", "responses", "limits", "germinal", "sizes"], "article_id"=>658520, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0060139.s001", "https://dx.doi.org/10.1371/journal.pone.0060139.s002", "https://dx.doi.org/10.1371/journal.pone.0060139.s003", "https://dx.doi.org/10.1371/journal.pone.0060139.s004", "https://dx.doi.org/10.1371/journal.pone.0060139.s005", "https://dx.doi.org/10.1371/journal.pone.0060139.s006"], "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Rgs13_Constrains_Early_B_Cell_Responses_and_Limits_Germinal_Center_Sizes__/658520", "title"=>"Rgs13 Constrains Early B Cell Responses and Limits Germinal Center Sizes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-03-23 02:26:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/995765"], "description"=>"<p>A. Flow cytometric analysis of WT and KI splenic B cell proliferation following stimulation with CD40 and IL-21 for 4 or 6 days. Proliferation assessed by Pacific Blue dye dilution. Representative results from the analysis of B cells from 3 WT and 3 KI mice. An analysis of GFP versus Pacific blue for the KI B cell is shown in the far left panels. B. Proliferative indexes from the flow cytometric analysis of WT and KI mouse spleen B cells stimulated as indicated. Results mean ± SEM of 3 WT versus 3 KI B cell preparations. Similar results from 2 other experiments using a partial overlapping set of inductive signals. C. Quantitative RT-PCR using RNA extracted from spleen cells from 10 day sRBC immunized KI/WT mixed chimera mice sorted for B220<sup>+</sup>CD38<sup>−</sup>GL7<sup>+</sup>CD95<sup>+</sup> and separated on the basis of CD45.1 or CD45.2. Results were normalized to <i>Gapdh</i> expression and expressed as ratio between KI and WT samples. Data are the mean ± SEM of triplicate values for each gene analyzed using data pooled from 3–4 separate experiments.</p>", "links"=>[], "tags"=>["proliferation", "abnormal", "ki", "gc"], "article_id"=>658514, "categories"=>["Immunology"], "users"=>["Il-Young Hwang", "Kyung-Sun Hwang", "Chung Park", "Kathleen A. Harrison", "John H. Kehrl"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0060139.g008", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Normal_B_proliferation_in_vitro_but_an_abnormal_gene_expression_pattern_in_Rgs13_GFP_KI_GC_B_cells_/658514", "title"=>"Normal B proliferation <i>in vitro</i>, but an abnormal gene expression pattern in <i>Rgs13</i>GFP KI GC B cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-23 02:22:40"}

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Relative Metric

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