Fig. 1. Nsun2 is essential for Th17 homeostasis.

a The protein level of Nsun2 in mouse organs were detected by western blot. b Nsun2 protein level in each T cell subsets was investigated by western blot. Sorted naive CD4⁺ T cells from lymph nodes of wild-type C57BL/6 mice were differentiated under Th1, Th2, Th17, and Treg inducing conditions respectively, and total proteins were then isolated from each T cell subsets. c, d FACS analyses and statistical (d) showing the percentage of CD4⁺ IL-17A⁺ Th17 cells in spleen and PBMC from littermate wild-type and Nsun2^−/− mice. Summary of the frequency of Th17 cells are shown in d (n = 8 for wild-type group and n = 7 for Nsun2^−/− group in spleen; n = 5 for wild-type and Nsun2^−/− groups in PBMC). e, f FACS (e) and statistical (f) analyses showing the in vitro differentiation frequency of Th17 cells. Sorted naive CD4⁺ T cells from wild-type and Nsun2^cKO mice respectively were induced under the Th17-inducing conditions. Th17 cells were in vitro differentiated for 4 days. Summary of the frequency of Th17 cells are shown in f (n = 9 per group). g The protein level of RoRγt in Th17 cells was detected by western blot. h, i FACS analysis showing the in vitro differentiation frequency of Th17 cells under digoxin treatment. Sorted naive CD4⁺ T cells from wild-type and Nsun2^cKO mice respectively were treated with DMSO control or digoxin (10 μM) before differentiated under Th17-inducing condition for 4 days. Summary of the frequency of Th17 cells are shown in i (n = 3 per group). All data are representative of at least two independent experiments. Data were analyzed using two-tailed unpaired Student’s t-test, Error bars represent mean ± s.e.m (d, f, i). Source data are provided as a Source Data file.