Figure 5.

IBP induces autophagy through downregulation of PAK1 in lung cancer cells

(A) Cells treated with the indicated concentrations of IBP (0, 5, 10, and 20 μg/mL) for 48 h. PAK1 expressions in A549 and H460 cells were measured by immunoblotting. (B) A549 cells were cotransfected with GFP-RFP-LC3 and Ctrl or PAK1 siRNA for 48 h and then were treated with 10 μg/mL IBP for another 48 h. CLSM was employed to determine LC3 puncta. Scale bar, 10 um. (C) A549 cells were cotransfected with GFP-RFP-LC3 and HA-PAK1 for 24 h, and then cells were treated as in (B). LC3 puncta were detected by CLSM. Scale bar, 10 um. (D) The expression of PAK1, p-Akt, and p-mTOR in xenograft tissues was assessed by immunoblotting. (E) In situ analysis of p-Akt and PAK1 abundance (brown) in xenograft tissues by IHC. Nuclei were counterstained by hematoxylin (blue). Scale bar, 50 μm. Data are mean ± SD from 3 independent experiments. One-way ANOVA (Tukey’s post hoc); ∗p < 0.05; ∗∗p < 0.01.