Fig. 3: PD-L1 inhibits apoptosis by promoting autophagy in EGFR-mutant LUAD cells. | Cell Death & Disease

Fig. 3: PD-L1 inhibits apoptosis by promoting autophagy in EGFR-mutant LUAD cells.

From: PD-L1 induces autophagy and primary resistance to EGFR–TKIs in EGFR-mutant lung adenocarcinoma via the MAPK signaling pathway

Fig. 3

A Autophagy was evaluated using TEM in PC9 PD-L1 and vector cells (left). Autophagosomes and autolysosomes were indicated by arrows (scale bar, 5 μm (upper) and 2 μm (lower)). The number of autophagosomes/autolysosomes was quantified (right). B Immunofluorescence assay revealing the effects of PD-L1 on PC9 and HCC827 cell autophagy (scale bar, 20 μm). C PC9 and HCC827 cells transfected with mRFP-GFP-LC3 lentivirus. PC9 PD-L1 and HCC827 PD-L1 cells were pretreated with 3-MA. Fluorescence microscopy was used to acquire images and the average number of yellow dots (autophagosomes) or red-only dots (autolysosomes) in the merged images per cell was quantified (scale bar, 20 μm). D Western blotting analysis of autophagy-related protein levels in PD-L1 and vector groups of PC9 and HCC827 cells. E. Apoptotic rate was measured using Annexin V/7-AAD staining in cells as in (C). F–H The viability of cells as in (C) was assessed by colony formation assay (F), EDU assay (G) and CCK-8 assay (H) (scale bar, 150 μm). I The apoptosis and autophagy-related protein levels were assessed by Western blotting in cells as in (C). The analyses were repeated three times. Data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

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