Fig. 4

The sensitivity of PKI-402 was increased and mtUPR was inhibited after inhibiting SGs. A After treatment with cycloheximide (CHX) 5 μg/ml for 45 min, washed twice with PBS, treated with PKI-402 (2.5 μM) for 12 h, and thapsigargin (Tg) 1 μM for 50 min. Colocalizations of YB-1 and G3BP1 was determined by staining and observed by fluorescence microscopy, scale bar, 10 µm. B Colocalizations of YB-1 and G3BP1 in C were quantified using ImageJ software. C After CHX treatment, PKI-402 (2.5 μM) was added for 12 h, and then cell viability was detected by MTT assays. D, F After CHX treatment, PKI-402 (2.5 μM) was added for 12 h, cell lysates were immunoblotted using the indicated antibodies. H Same treatment with D, F, then mitochondrial proteins were collected and the expression of mitochondrial-associated protease and chaperone was analyzed by Western blotting. E, G, I Levels of the proteins in D, F, and H were measured by Western blotting, respectively. Data were expressed as mean ± SEM of three independent experiments; **P < 0.01, ****P < 0.0001 compared to control. J, K After CHX treatment, PKI-402 (2.5 μM) was added for 12 h, and A2780 cells were treated with PKI-402 (2.5 μM) for 12 h and then incubated with DCFH-DA and JC-1. Changes in fluorescence intensity were measured by flow cytometry