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. 2011 Feb 15;25(4):310–322. doi: 10.1101/gad.1984311

Figure 5.

Figure 5.

Effect of RNAi-mediated knockdown of FoxO1 on autophagy and survival of JNKTKO neurons. (A,B) Wild-type (control) and Jnk1LoxP/LoxP Jnk2−/− Jnk3−/− (JNKTKO) neurons infected with Ad-cre at 3 DIV were transfected at 7 DIV with FoxO1 siRNA or control siRNA. The expression of FoxO1 mRNA (A) and Bnip3 mRNA (B) was examined at 11 DIV by quantitative RT–PCR analysis of mRNA and normalized to the amount of Gapdh mRNA in each sample (mean ± SD; n = 3). Statistically significant differences are indicated. (*) P < 0.05. (C) Control and JNKTKO neurons transfected with scrambled sequence or FoxO1 siRNA were examined at 11 DIV by immunoblot analysis with antibodies to LC3b, p62/SQSTM1, and α-Tubulin. (D) RNAi transfected JNKTKO neurons were examined at 11 DIV by quantitative RT–PCR analysis of Atg3, Atg5, and Atg12 mRNA and normalized to the amount of Gapdh mRNA in each sample (mean ± SD; n = 3). Statistically significant differences are indicated. (*) P < 0.05. (E) The survival of RNAi transfected control and JNKTKO neurons at 11 DIV was quantitated (mean ± SD; n = 20). Statistically significant differences are indicated. (*) P < 0.05.