Extended Data Fig. 5: Gene expression in inertially cycling B cells.
From: Transient silencing of hypermutation preserves B cell affinity during clonal bursting
The anti-DEC-OVA-treated and control GC B cells analysed for mutational load in Fig. 2g–j were profiled for expression of genes involved in SHM. a, Expression of Aicda and downstream DNA repair genes in GC B cells prior to and at 48 and 72 h post-treatment. P values are given for the comparison between treated cells at 48 h post-anti-DEC-OVA (not actively undergoing SHM) and 72 h post-anti-DEC-OVA (actively mutating). Each symbol represents the mean expression value of single B cells from one mouse. Data are from two independent experiments. Expression of Ung, Apex1, Apex2, Neil1, Neil3, and Xrcc4 (involved in base excision repair); Msh2 and Msh6 (required for mismatch repair); and the error-prone polymerase Polh, Hmces, and Fam72a (involved in the error-prone processing of AID-induced lesions) was not lower at 48 h. b, Expression of an AID-GFP fusion reporter analysed by flow cytometry at different times after anti-DEC-OVA treatment. Left, representative histograms; right, quantification of 2–4 samples for each time point. Each symbol represents one mouse. c, Detection of Vh1-72 mRNA (encoding for the heavy chain of the B1-8hi B cell receptor) and of the intronic RNA downstream of all Ighj segments (Jh4 intron, indicative of active Igh transcription) in GC B cells prior to and at 48 and 72 h post-anti-DEC-OVA treatment. Each symbol represents the mean expression value of single B cells from one mouse. Data are from two independent experiments. d, Gene-expression-based cell-cycle assignments for GC B cells from each time point (data as in a,b). Cells are pooled from 7–9 mice from 2 independent experiments as in a,c. Cell-cycle categories are assigned using the CellCycleScoring function in Seurat. G0/G1 cells are defined as those lacking expression of S-phase and G2/M-phase transcripts.
