Fig. 8

Regulation by C/EBPβ and NFIC silencing of PD-L1 expression in NSCLC cell lines carrying different rs822336 genotype under basal conditions or following IFN-ɣ incubation. EGFR^mut H1975^G/G and EGFR^wt H460^G/G cells (upper panel) as well as EGFR^wt H1299^C/C and EGFR^wt H1437^C/C cells (lower panel), transduced with C/EBPβ- and NFIC-specific siRNAs or siRNA-controls, were seeded into 6-well plates at a density of 2 × 10⁶ cells per well and incubated with IFN-ɣ (100ng/ml). Untreated cells were used as a control. Following a 48 h incubation at 37 °C in a 5% CO₂ atmosphere, cells were harvested and lysed. Cell lysates were analyzed by western blot with NFIC, C/EBPβ and PD-L1-specific Abs. Representative results are shown. The levels of NFIC and C/EBPβ, normalized to GAPDH and relative to untreated siRNA-control as well as levels of PD-L1, normalized to GAPDH and relative to treated siRNA-control, are plotted on the right, and expressed as mean ± SD of the results obtained in three independent experiments