Skip to main content
. 2019 Aug 30;294(42):15435–15445. doi: 10.1074/jbc.RA119.009618

Figure 5.

Figure 5.

Brain-specific somatic mutation in Akt3 debilitates Siah1-mediated Akt3 degradation. A, cells were transfected with GFP-Siah1 and FLAG-Akt-WT or E17K. After 24-h transfection, the cell lysates were subjected to an immunoprecipitation (IP) assay. The total amount of protein was checked with anti-actin antibody. B, PC12 cells were transfected with the indicated amounts of Myc-Siah1. The level of Akt3-WT or E17K was detected by immunoblot. The graph shows the percentage of Akt3-WT and E17K protein levels (right panel). Data are shown as mean ± S.E. of three independent experiments; one way analysis of variance. ****, p < 0.0001. C, PC12 cells were transfected with the indicated constructs. After transfection for 24 h, cells were treated with 10 μm MG132 for 8 h. The cell lysates were immunoprecipitated with the anti-FLAG antibody. The ubiquitination was checked with the anti-HA antibody. D, the hippocampal neurons were transfected with the indicated constructs at DIV 3 and fixed at DIV 5. Data are shown as mean ± S.E. of three independent experiments. *, p < 0.05 versus control; **, p < 0.005 versus control; #, p < 0.05. The images shown here are representative of at least three independent experiments.