Skip to main content
. 2021 Aug 4;2021:9937243. doi: 10.1155/2021/9937243

Figure 5.

Figure 5

HuR deficiency increases IL-22 production in Th17 cells by reducing c-Maf expression. Naïve CD4+ T cells from WT and HuR KO mice were cultured under Th17 cell polarization condition for 5 days. (a) The expression of c-Maf and IL-22 mRNAs was examined by RT-qPCR. IL-22 mRNA RT-qPCR is a positive control in (a). (b) The levels of IL-22 in WT and HuR KO Th17 cell culture supernatant were examined by ELISA. (c) The abundance of c-Maf and HuR protein in WT and HuR KO Th17 cells was examined by western blots. (d) Naive WT CD4+ T cells were transduced with c-Maf siRNA and scramble siRNA and polarized under Th17 cell culture condition for 5 days. The expression of IL-22 and c-Maf mRNA was measured by RT-qPCR assay. (e) HuR bound to c-Maf mRNA as determined by RIP assay; one of the two repeated experiments is shown. The repeated RIP experiment got the same tendency of results. (f) Actinomycin D (3 μg/ml)-treated Th17 cells were harvested at 1, 3, and 5 h; the level of c-Maf mRNA was determined by RT-qPCR assay. One of the two repeated experiments is shown. (g) Polysome fraction assay showed that HuR did not modulate c-Maf mRNA translation. Results in (a) and (b) represent the summary of three independent experiments (mean ± SEM). Student's t-test was used for statistics analysis. ∗∗p < 0.01 and ∗∗∗p < 0.001. Data in (c)–(g) represent one of the two independent experiments.