Figure 3.
SOD1 silencing in SOD1G93A iPSA decreases MTOR activity. CTRL and G93A iPSA were transduced with shSOD1 lentiviral particles (+) or negative control (-) for 48 h prior to the assays. (A) SOD1 mRNA levels normalized to ACTB (n = 3). (B) Representative western blot and quantification of SOD1 protein levels normalized to ACTB (n = 3), (C) p-MTOR (Ser2448) protein level normalized to total MTOR (n = 3), (D) p-EIF4EBP1 (Thr37/46) protein level normalized to total EIF4EBP1 (n = 3). (E) Representative images of CTRL and G93A iPSA stained with BrdU (green) and DAPI (blue), and quantification of cell proliferation (n = 7). (F) HSBP1, GFAP, SERPING1, GGTA1, GBP2 mRNA levels normalized to ACTB (n = 3). P values <0.05 by one-way ANOVA test with Sidak’s correction between indicated groups are shown