Fig. 6

Rolapitant triggered ER stress through down-regulating GRP78 and upregulated CHOP-DR5 pathway. A Western blot analysis of PERK, p-PERK, EIF2α, p-EIF2α, ATF4, ATF6, IRE1, XBP-1 s and CHOP levels in A549 and H1299 cells treated with DMSO or indicated concentrations (10, 15, 20, or 25 μM) of Rolapitant for 48 h. GAPDH was used as a loading control. B Western blot analysis of ATF4, CHOP, DR5, Caspase-8, Cleaved-Caspase-8, Caspase-3, Cleaved-Caspase-3, PARP and Cleaved-PARP levels in A549 and H1299 cells treated with DMSO or Rolapitant (10 μM) in the absence or presence of TM (1 μg/ml) for 24 h. C Western blot analysis of GRP78, CHOP, DR5, PARP, Cleaved-PARP and OTUD3 in OTUD3 shRNA knocking down and control A549 and H1299 cells. GAPDH was used as a loading control. D, E Western blot analysis of CHOP, DR5, Caspase-8, Cleaved-Caspase-8, Caspase-3, Cleaved-Caspase-3, PARP, Cleaved-PARP and OTUD3 in OTUD3 shRNA knocking down and control A549 and H1299 cells treated with or without Rolapitant (10 μM) for 48 h. GAPDH was used as a loading control