Fig. 2: RIF1 limits the ex vivo differentiation of activated B cells to PBs and PC-like cells independently from its role in DNA end protection and CSR. | Nature Communications

Fig. 2: RIF1 limits the ex vivo differentiation of activated B cells to PBs and PC-like cells independently from its role in DNA end protection and CSR.

From: RIF1 integrates DNA repair and transcriptional requirements during the establishment of humoral immune responses

Fig. 2

a Left: Representative flow cytometry plots measuring percentage of plasmablasts (CD138 + ) in purified splenic B cell cultures 96 h after activation with LPS and IL-4. Right: Summary graph for five independent experiments (n = eight mice per genotype), with values within each experiment normalized to the average of controls, which was set to 100%. b Left: Representative plots measuring proliferation by CellTrace Violet (CTV) dilution and percentage of CD138+ cells per cell division. Right: Summary graph for three independent experiments (n = six mice per genotype). c Left: Representative plots measuring percentage of CD138+ cells and summary graph for four independent experiments (n = thirteen (control) and seven mice (for the respective genotypes)). Right: Graph summarizing percentage of IgG1+ cells in the same cultures. CD138+ and CSR values within each experiment were normalized to the average of control mice (WT and Cd19Cre/+: Ctrl), which was set to 100% (n = thirteen (control) and seven mice (for the respective genotypes)). d Schematic representation of the iGB cell culture system for the ex vivo differentiation of splenic naïve B cells to GC- and plasma cell (PC) -like cells. CD40L: CD40 ligand; d: day; Sp: splenic. Left: Representative plots measuring CSR to IgG1 (e) and percentage of plasmablasts (f) at day 4 (e) and 8 (f), respectively, of the iGB cell culture. Right: Summary graphs for five independent experiments (n = eight mice per genotype). g Graphs depicting the percentage of IgM+ and IgG1+ cells in the CD138+ populations at the indicated timepoints of the iGB cell culture. n = six Cd19Cre/+ mice for IgM, five Rif1F/FCd19Cre/+ for IgM, five Cd19Cre/+ for IgG1, and four Rif1F/FCd19Cre/+ for IgG1 in three independent experiments. Significance in panels (a, b, c, e, f, g) was calculated with a two-sided Mann–Whitney U test, with data presented as mean values ± SD in panels (a, c, e, f, and g), and SEM in panel (b). Only significant differences were indicated in panel (b). ns: not significant. Source data are provided as a Source Data file. Created in BioRender. Di virgilio, M. (2025) BioRender.com/f53q006.

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