Cultured neurons were cotransfected with the indicated knockdown and expression constructs at 12 DIV and subjected to AHA labeling for 1 h at 18 DIV. (a,c,e) Representative images of AHA labeling. The insets indicate the transfected cells, which are outlined or indicated by arrows in the original images. (b,d,f) The relative AHA intensity in somata. In (e), AHA omission or cycloheximide addition (Chx; 10 μM) was used to confirm the specificity of the AHA labeling. To indicate the presence of neurons, DAPI signals are shown in these two groups. Ctrl, non-silencing control. Scale bar: 25 μm. The sample sizes (n) of the examined neurons are indicated. The data from three independent experiments are presented as the mean+s.e.m. (error bars). *P<0.05; **P<0.01; ***P<0.001. One-way ANOVA (b,d), two-way ANOVA (f).