Figure 4.

Effect of RNAi-mediated knockdown of Beclin-1 on autophagy and survival of JNK^TKO neurons. (A) Wild-type (control) and Jnk1^LoxP/LoxP Jnk2^−/− Jnk3^−/− (JNK^TKO) neurons infected with Ad-cre at 3 DIV were transfected at 7 DIV with Beclin-1 siRNA or control siRNA. The expression of Beclin-1 mRNA 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. (B) Control and JNK^TKO neurons transfected with scrambled sequence or Beclin-1 siRNA were examined at 11 DIV by immunoblot analysis with antibodies to LC3b, p62/SQSTM1, and α-Tubulin. (C) The survival of RNAi transfected control and JNK^TKO neurons at 11 DIV was quantitated (mean ± SD; n = 20). Statistically significant differences are indicated. (*) P < 0.05.