Fig. 2

TGF-β1-induced EMT increases PD-L1 expression in H358 cells. Cells were treated with TGF-β1 (10 ng/ml) for 48 h. a PD-L1 mRNA expression was measured by RT-qPCR and normalized to GAPDH mRNA. The graph corresponds to the mean ± SD of three independent experiments. ***p < 0.001 by Student’s t test. b PD-L1 expression was determined by immunoblotting of cell lysates. Actin was used as a loading control. The apparent molecular weights (kDa) are indicated on the right of the panel. c PD-L1 (green) was detected by immunofluorescence and analyzed by confocal microscopy. DNA (red) was stained with DRAQ5. d Representative flow cytometry experiment of PD-L1 staining in H358 cells. Histograms represent the mean ± SD of PD-L1 MFI (mean fluorescence intensity) of three independent experiments (right panel). *p = 0.0391 by Student’s t test. NT no TGF-β1 treatment