Fig. 3: Nuclear TFEB reflects antigenic B cell experience.

a–f Splenocytes of aged-matched wild-type C57BL/6 mice were left untreated or stimulated with anti-IgM and anti-IgG for 60 min. Among splenic CD19⁺ B cells, CD80^- B cells and CD80⁺ memory B cells were differentiated through surface staining (a). TFEB nuclear translocation was analyzed by imaging flow cytometry as described in Fig. 1. b Representative multi-channel images of individual cells. c Histograms depicting TFEB/7-AAD similarity scores of CD80^- and CD80⁺ B cells in the absence of BCR ligation. d Percentages of cells with similarity scores of TFEB/7-AAD ≥ 1 in resting and BCR-stimulated subsets. e, f Expression of TFEB (e) within the whole cell and (f) within the nucleus. g–j Primary human B cells were isolated from the blood of healthy donors and left untreated or BCR-stimulated for 60 min. Localization of TFEB in individual primary human B cell subpopulations is defined by the following cell surface staining patterns. CD19⁺CD27^-IgD⁺ (naïve), CD19⁺CD27^- (total non-memory), CD19⁺CD27⁺ (total-memory), CD19⁺CD27⁺IgD⁺ (unswitched memory), CD19⁺CD27⁺IgD^- (switched memory), or CD19⁺CD27⁺IgD^-CD38⁺ (plasmablasts). The corresponding gating strategy is shown in Supplementary Fig. 2f. g Representative multi-channel images of individual cells. h Histogram analysis of naïve B cells, or unswitched and switched memory B cells in the absence of BCR ligation. of TFEB (FITC) versus 7-AAD similarity score. i Percentages of cells with similarity scores of TFEB/7-AAD ≥ 1 in resting and BCR-stimulated B cell subsets. j TFEB expression among resting B cell subsets. All data are depicted as mean ± SD of n = 3 independent experiments. Statistical significances were calculated using one-way ANOVA and corrected for multiple testing via Tukey’s method. Source data are provided as a Source Data file.