Rab1A-knockdown-induced cell death was caused by inhibition of autophagy but not ER-Golgi traffic.

<p>(A) BFA induced cell death differently from Rab1A knockdown. HT29 cells were transfected with <i>Hsc70</i> or <i>control</i> siRNA and untransfected cells were treated with 5 µg/ml BFA or DMSO at the onset of IncuCyte cell growth assay. Images were captured to monitor proliferation by IncuCyte inside a cell culture incubator. (B) Phase-contrast images at 48 h after the onset of measurement. Scale bar, 200 µm. (C) Inhibition of autophagy was not caused by the interruption of ER-Golgi traffic. Rab1A knockdown or control cells were subjected to serum depletion, 5-FU, or vehicle treatment for 24 h. Untransfected cells were treated with 5 µg/mL BFA for 6 h, and immunoblotted with anti-p62, LC3B, or Rab1A. β-actin was used as a loading control. (D) Hsc70 knockdown allowed autophagosome formation. HT29 cells transfected with <i>Hsc70</i>, <i>Rab1A</i>, <i>Ran</i>, or <i>control</i> siRNA were subjected to serum depletion or vehicle treatment for 24 h, and immunoblotted. Immunoblotting data are representative of at least two separate experiments yielding similar results.