Extended Data Fig. 3: Arid1a deficiency does not have a major impact of B cell activation in vitro. | Nature Immunology

Extended Data Fig. 3: Arid1a deficiency does not have a major impact of B cell activation in vitro.

From: Arid1a-dependent canonical BAF complex suppresses inflammatory programs to drive efficient germinal center B cell responses

Extended Data Fig. 3

a. Representative flow cytometry analysis showing YFP expression in Cg1 Arid1a KO B cells at pre- and post-stimulation (day 4) on 40LB co-cultures with IL4 (in vitro). b. Representative flow cytometry analysis showing GL7 expression in Cg1cre (controls) and Cg1 Arid1a KO B cells at day 4 of in vitro 40LB co-culture. c. Representative flow cytometry analysis of IgD and IgM expression from Arid1a fl/fl (YFP-) and CD19 Arid1a KO (YFP+) splenic B cells. The median fluorescence intensity (MFI) for IgD in quadrant1 (Q1) is shown in the plots. d. Representative flow cytometry analysis quantifying percent apoptotic cells (cleaved Caspase3) from Cg1 cre (n = 4) and Cg1 Arid1a KO (n = 3) B cells at day 4 in vitro 40LB co-cultures. e. Quantification of apoptotic B cells (cleaved Caspase 3 positive) from Cg1 cre (n = 4) and Cg1 Arid1a KO (n = 3) on 40LB co-cultures. f. Representative flow cytometry analysis of proliferating cells (Ki67 positive) from Cg1 cre and Cg1 Arid1a KO B cells at day 4 of in vitro 40LB co-cultures. g. Quantification of proliferating B cells (Ki67 positive) from Cg1 cre (n = 4) and Cg1 Arid1a KO (n = 3) on 40LB co-cultures. h. Representative flow cytometry analysis quantifying percent plasma cells (CD138+) from Cg1 cre and Cg1 Arid1a KO B cells at day 4 of in vitro 40LB co-cultures. i. Quantification of percent of CD138+ plasma cells from Cg1 cre (n = 4) and Cg1 Arid1a KO (n = 3) B cells at day 4 of in vitro 40LB co-cultures. j. Representative flow cytometry analysis of IgG1 and IgM positive B cells from Arid1a fl/fl and CD19 Arid1a KO mice after 4 days of 40 LB co-cultures. k-l Quantification of IgM and IgG1 frequencies from day 4 of 40LB co-culture (n = 3). m-n. Median fluorescence intensity (MFI) of IgM and IgG1 from day 4 of 40LB co-culture (n = 3). o. Cell counts of Arid1a fl/fl and CD19 Arid1aKO B cells stimulated on 40LB co-cultures for 4 days (n = 3). p-q Quantification of apoptotic (cleaved Caspase 3 positive) and proliferating B cells (Ki67 positive) respectively from Arid1a fl/fl and Cd19 Arid1a KO on 40LB co-cultures (n = 3). r. Representative flow cytometry analysis quantifying percent plasma cells (CD138+) from Arid1a fl/fl and Cd19 Arid1a KO B cells at day 4 of in vitro 40LB co-cultures. s. Quantification of percent of CD138+ plasma cells from Arid1a fl/fl and Cd19 Arid1a KO B cells at day 4 of in vitro 40LB co-cultures (n = 3). Statistical significance is calculated using two-tailed Student’s t-test for e, g, i, k-q, and s. Error bars represent mean ± s.e.; *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.0005; ****p value < 0.0001.

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