DDIT3 positively regulates the B cell activation and BCR signaling. Splenic B cells from WT and DDIT3 KO mice were stimulated with anti-F(ab’)2 goat anti-mouse IgG + IgM (10 μg/mL) for the indicated time, and the BCR signaling was analyzed by confocal and WB. (A–C) Confocal analysis of DDIT3 in splenic B cells from WT mice. Representative pictures and Pearson correlation coefficients between BCR and DDIT3/DAPI. (D–F) Confocal analysis of pY and pBLNK in splenic B cells from WT and DDIT3 KO mice. Representative plots and Pearson correlation coefficients between BCR and pY, BCR and pBLNK. Scale bar, 2 μm. Data were collected from more than 50 cells. (G, H) Immunoblot of pY, pBLNK, pSyk, pPI3K, pAkt1, pmTOR, pFoxO1, pS6, and total proteins in B cells from WT and DDIT3 KO mice. Representative plots from three independent experiments are shown. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001, unpaired student’s t-test and one-way ANOVA.
