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. 2024 Nov 23;15:10163. doi: 10.1038/s41467-024-54344-5

Fig. 7. Rag-GTPases-TFEB/TFE3 axis modulates humoral immunity in a context dependent manner.

Fig. 7

A Flow cytometry of CD43 and B220 expression on BM cells. Below, summaries of B220loCD43−, B220hiCD43− and B220+CD43+ cell frequencies. B Flow cytometry of BP-1 and CD24 expression in BM B220+CD43+IgM– B cell precursors. Right, summary of fraction A (CD24−BP-1−), fraction B (CD24+BP-1−), and fraction C/C′ (CD24+BP-1+) cell frequencies. C Flow cytometry of GL-7 and Fas expression the Peyer’s patches. Right, summary of the GC B cell frequencies. For A-C, WT (n = 5), μMT:CreERRragafl/flRragbfl/fl (n = 6), and μMT:CreERRragafl/flRragbfl/flTfebfl/flTfe3−/− (n = 6). Splenic B cells were stimulated with LPS/IL-4/BAFF for 72 h. IgG1 expression (D), TMRM and MTDR staining (E), CellROX staining (F), and LAMP1 staining (G) were examined by flow cytometry. Summaries of IgG1+ B cell frequencies, the relative TMRM, MTDR, CellROX, and LAMP1 MFIs were presented. For (D–G), n = 4 mice for each group. H Mitostress assay was performed on a Seahorse XFe96 analyzer. Right, summaries of the basal respiration and maximal respiration. WT (n = 6), μMT:CreERRragafl/flRragbfl/fl (n = 5), μMT:CreERRragafl/flRragbfl/flTfebfl/flTfe3−/− (n = 6). I–L Tamoxifen injected WT, μMT:CreERRragafl/flRragbfl/fl, and μMT:CreERRragafl/flRragbfl/flTfebfl/flTfe3−/− chimera mice were immunized intraperitoneally with NP-OVA/alum (I, J) or TNP-LPS (K, L). I Flow cytometry of CD138 and B220 expression on splenic lymphocytes. Right, summary of CD138+ plasmablast frequencies. J Flow cytometry of GL-7 and Fas expression on splenic B cells. Right, summary of GC B cell frequencies. For (I and J), WT (n = 4), μMT:CreERRragafl/flRragbfl/fl(n = 3), and μMT:CreERRragafl/flRragbfl/flTfebfl/flTfe3−/− (n = 4). K Flow cytometry of CD138 and B220 expression on splenic lymphocytes. Right, summary of CD138+ plasmablast frequencies. L ELISA measurement of serum TNP-specific antibodies. For (K and L), WT (n = 6), μMT:CreERRragafl/flRragbfl/fl (n = 7), μMT:CreERRragafl/flRragbfl/flTfebfl/flTfe3−/− (n = 6). Data in graphs represent mean ± SEM. ns, not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001, one-way ANOVA (A, C, D–K), two-way ANOVA (B and L). Source data are provided as a Source Data file.