Fig. 5
XPO1 inhibitor KPT-330 induced autophagy of GBC cells through the mTOR pathway. A Transmission electron microscopy in NOZ and GBC-SD showed more autophagosomes and autolysosomes after KPT-330 treatment compared with DMSO treatment group. Red arrows point to autophagosomes and autolysosomes. Scale bars represent 1.0 μm. B mCherry-GFP-LC3 dual fluorescent images indicated that green, red, and yellow (from the merged images) puncta increased greatly in KPT-330 treated cells compared to DMSO treatment in NOZ and GBC-SD. Scale bars represent 100 μm. C Expression of LC3- II/I and p-mTOR/mTOR was detected by western blot after KPT-330 treatment for 48 h. D Expression of LC3- II/I was detected by western blot after KPT-330 treatment at 0, 12, 24, 48 h. E Expression of LC3- II/I and p-mTOR/mTOR was detected by western blot after KPT-330 or rapamycin or chloroquine treatment. GBC cells were pre-treated with chloroquine (concentration of 0.2 μM) or rapamycin (concentration of 0.4 μM) for 6 h, then were treated with or without KPT-330 for 48 h. F Chloroquine attenuated inhibition effects of KPT-330 in NOZ and GBC-SD assessed by cell proliferation assays. GBC cells were pre-treated with chloroquine (concentration of 0.2 μM) for 6 h, then were treated with or without KPT-330 for 48 h. Student's t test was applied to the statistical analysis in this figure. Data presented as mean ± SD (n = 3)