Fig. 4.

Effect of inhibition of GRP78 on cell death pathways in MIA PaCa-2 and S2-VP10 cells. A–C: MIA PaCa-2 and S2-VP10 cells were treated with GRP78 siRNA pool and assayed for caspase-3 activation at 48 and 72 h posttransfection (A and B) and annexin V staining at 48 h posttransfection (C) and compared with cells treated with transfection reagent alone (control) or with nonsilencing siRNA. D–F: autophagy was assayed by monitoring LC3B levels by Western blotting (D) or by immunofluorescence (E) and quantitated (F). Data are representative of 3 independent experiments. G and H: autophagy was inhibited by knockdown of BECLIN1 and/or GRP78, and the effect on autophagy was monitored by assaying for LC3B by immunofluorescence (G) and shown as cell viability (H) compared with cells treated with nonsilencing siRNA or the GRP78 siRNA pool alone. Results are representative of 4 independent experiments. Values are means ± SE; n = 3. *P < 0.01.