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. Author manuscript; available in PMC: 2020 Dec 22.
Published in final edited form as: Nat Immunol. 2020 Jun 22;21(7):777–789. doi: 10.1038/s41590-020-0706-5

Extended Data Fig. 5. ATAC-seq of TFH and non-TFH populations from multiple genetic backgrounds.

Extended Data Fig. 5

Related to Fig. 5

a, Representative flow cytometry of naive SMARTA cells (CD44loCD62LhiCD45.1+) used for ATAC-seq analysis.

b, Genome-browser tracks depict ATAC-seq chromatin accessibility at Cxcr5 locus. Peak calls indicated below each track. * indicates DEseq2 raw p ≤ 0.05 in comparison between WT TFH and TH1.

c, Genome-browser tracks depict ATAC-seq chromatin accessibility at Tbx21 and Gata3 loci.

Peak calls indicated below each track. Bcl-6 liftover peaks from human to mouse reference genome are indicated. * and ** indicate DEseq2 raw p-val 0.05 and 0.01, respectively, in comparison between WT TFH and TH1. Gene expressions from RNA-seq data of LCMVArm infected mice are graphed. DKO, Bcl6f/fPrdm1f/f CreCD4.

d, Genome-browser tracks depict BCL-6 ChIP-Seq peaks at SELPLG locus. Peak calls indicated below the track.

e, Representative flow cytometry and quantification of the level of Bcl-6 in RV+ TFH or non-TFH SMARTA cell subsets from spleen of LCMVArm infected mice. SMARTA CD4 T cells transduced with GFP-RV or Bcl6-RV were transferred to C57BL/6 host mice, followed by infection of the host mice with LCMVArm, and analyzed 3 days after infection. Non-TFH: GFP-RV+ and TFH: GFP-RV+ indicate the expression level of endogenous Bcl-6 in each population. Each dot represents one mouse (n = 4). Data are mean s.d., unpaired two-tailed Student’s t-test.

f, Representative flow cytometry and quantification of GC-TFH and BGC subpopulations from spleen of LCMVArm infected mice. Bcl6f/f CreCD4 SMARTA CD4 T cells transduced with Bcl6-RV or myctagN-Bcl6-RV were transferred to Bcl6f/f CreCD4 host mice, followed by infection of the host mice with LCMVArm, and analyzed 8 days after infection. We validated that an myctagN-Bcl6-RV was functionally comparable with non-tagged Bcl6-RV (Bcl6-RV), based on myctagN-Bcl6-RV rescue of TFH differentiation and B cell help function of Bcl6f/f CreCD4 CD4 T cells in LCMV infected mice. Each dot represents one mouse (n = 4). Data are mean s.d.