Fig. 5: Increased frequency of generation of pathogenic Th17 cells in culture in the absence of Senp2.

a, b FACS showed the production of pathogenic Th1 and Th17 cells by differentiating naive CD4⁺ T cells from WT and CKO mice for 5 days (a). Statistical analysis is shown in (b). c Genomic PCR showing deletion of the Senp2 allele in the presence of 500 nM 4-OHT in differentiating pathogenic Th17 cultures derived from CKO-ER (Senp2^f/f ERcre⁺) mice at the indicated days, compared with those from WT-ER (Senp2^f/f ERcre⁻) mice. P-Selectin was used as an internal control. d, e FACS showing the frequency of IFNγ⁺ (top) and IL-17A⁺ (bottom) cells at day 5 of pathogenic Th17 polarizing culture derived from CKO-ER⁺ and WT mice with the addition of 4-OHT (d). Statistical analysis is shown in (e). f–h Re-introduction of wild-type SENP2 (Se) or catalytic domain mutant of SNEP2 (c/s) via GFP carrying lentiviral vector into differentiating pathogenic Th17 culture derived from either WT or CKO mice. Immunoblotting showing the expression of exogenous SENP2 in CKO culture transduced with vector alone (Ctrl), wild-type SENP2 (Se.), or c/s mutant of SENP2 at 48 h after transduction (f). FACS showing the frequency of IL-17A⁺ cells after re-introduction of Ctrl, wild-type SENP2 (Se.), or c/s SENP2 in differentiating pathogenic Th17 culture derived from either WT or CKO mice (g). Statistical analysis is shown in (h). Results in (b, e, h) are mean ± SD (n = 3). Statistical analysis was done by Student’s t test. *P < 0.05. ns. not significant.