1. FIP200 maintains microRNA1198-5p expression via Ago2 in naïve T cells.
FIP200 maintains microRNA1198-5p expression via Ago2 in naïve T cells. (A) Expression of Ago2 protein in WT and Fip200−/− naïve T cells (n = 3). (B) Effect of Ago2 siRNAs on microRNA1198-5p expression. WT naïve CD4+ T cells were transfected with Ago2 siRNAs (#1 and #2) for 48 hours. The expression of micorRNA1198-5p and microRNA342-5p was quantified by real-time PCR (n = 3 to 5, means ± SEM). *P < 0.05 (Student’s t tests). (C and D) Expression of microRNA1198-5p (C) and Bak1 mRNA (D) in the Ago2 complex (n = 3, means ± SEM). *P < 0.05 (Student’s t tests). (E) Contents of Bak1 and Bax mRNAs in the Ago2 complex in WT and Fip200−/− naïve T cells. The data are shown as the percentage of Bak1 and Bax mRNA expression compared with input (n = 3, means ± SEM). *P < 0.05 of Bak1 expression between WT and Fip200−/− T cells (Mann-Whitney U tests). (F) Ago2 ubiquitination in WT and Fip200−/− naïve T cells. Naïve T cells were treated with or without proteasome inhibitor MG132 (10 μM) for 2 hours (n = 3). IP, immunoprecipitation; IB, immunoblotting. (G) Expression of Ago2 and Bak protein in Atg3-WT and Atg3-KO naïve T cells (n = 3).
Houjun Xia et al. Sci. Immunol. 2017;2:eaan4631
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