TP421 affects the autophagic response in pancreatic cancer cells.

(A) BxPC-3 and MIA PaCa-2 cells were treated with 0.5 and 2.5 µM TP421 or chloroquine for 24 and 48 h and probed for Beclin1, p62, and LC3B-I/II. (B) LC3B puncta formation in MIA PaCa-2 cells treated with TP421 for 18 h alone or in combination with rapamycin or chloroquine. (C) MEF cells competent in autophagy (atg3+/+) and deficient in autophagy (atg3−/−) were treated with increasing concentrations of TP421 and percent cytotoxicity was determined by MTT. The data are mean ± SD from three independent experiments. * and ** indicate p<0.05 and p<0.001 respectively. (D) MEF atg3+/+ and −/− cells treated with TP421 for 24 h and probed for autophagy proteins LC3B-I/II and p62. (E) MIA PaCa-2 cells treated with TP421 for increasing durations of time and co-stained with LysoTracker Red (LTR).