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. 2020 Oct 22;8:553728. doi: 10.3389/fcell.2020.553728

FIGURE 5.

FIGURE 5

Six3os1 activates the Fezf1/AKT axis by binding to miR-511-3p, thus ameliorating oxidative stress of CUMS-induced neurons. (A) Fezf1 mRNA expression in neurons after alteration of Fezf1 detected by RT-qPCR. (B,C) Immunoblots (B) and quantitation (C) of Fezf1 protein expression normalized to GAPDH in neurons after alteration of Fezf1 examined by Western blot analysis. The CUMS-induced neurons were treated with oe-NC + sh-NC, oe-Fezf1 + sh-NC, oe-Six3os1 + sh-Fezf1, miR-511-3p inhibitor + sh-Fezf1, oe-Six3os1 + sh-NC or miR-511-3p inhibitor + sh-NC, and CUMS-induced neurons with GP treatment were infected with oe-NC + sh-NC, oe-NC + sh-Fezf1, oe-NC + sh-Six3os1, oe-Fezf1 + sh-Six3os1, miR-511-3p mimic + oe-NC or miR-511-3p mimic + oe-Fezf1. (D–G) SOD (D), GSH (E), CAT (F), and MDA (G) contents in neurons determined by ELISA. (H–K) Quantitation of Bax (H), Cleaved-caspase3 (I), Bcl-2 (J), and caspase3 (K) protein expression normalized to GAPDH in neurons detected by Western blot analysis. (L–N) Immunoblots (L) and quantitation of AKT expression (M) and p-AKT (N) normalized to GAPDH in neurons determined by Western blot analysis. The measurement data were presented as mean ± standard deviation. Data between two groups were compared by independent sample t-test, and data among multiple groups were analyzed by one-way ANOVA, followed by Tukey’s post hoc test. Cell experiments were repeated three times. ∗p < 0.05 vs. oe-NC + sh-NC, oe-Six3os1 + sh-NC, miR-511-3p inhibitor + sh-NC, oe-NC + sh-Six3os1 or miR-511-3p mimic + oe-NC.