Fig. 2

SCG2 OE influences Stat1-induced MHC class I surface presentation, which can be partially restored by IFNγ treatment (A) Mean fluorescence intensity (MFI) of HLA-ABC-positive ( +) WM266-4 (left panel) and C32 (right panel) EV and SCG2 OE cells before and after IFNγ treatment (10 ng/ml, 48 h). Data represent mean ± s.e.m. (n ≥ 3). B Fold change of Stat1 mRNA expression in WM266-4 (left panel) and C32 (right panel) EV and SCG2 OE cells before and after IFNγ treatment (10 ng/ml, 48 h). 18S was used as endogenous control. Data represent mean ± s.e.m. (n ≥ 3). C Western blot analysis of the expression of total Stat1 and pStat1 (phosphorylated Stat1) in WM266-4 (left panel) and C32 (right panel) EV and SCG2 OE cells before and after IFNγ treatment (10 ng/ml, 48 h). GAPDH was used as a loading control. D Fold change of mRNA expression levels of the APM components TAP1, TAP2, B2M, and TAPBP (tapasin) in WM266-4 (left panel) and C32 (right panel) EV and SCG2 OE cells before and after IFNγ treatment (10 ng/ml, 48 h). 18S was used as an endogenous control. Data represent mean ± s.e.m. (n ≥ 3). E Western blot analysis of the expression of the APM components TAP2, TAP1, tapasin, and B2M in WM266-4 (left panel) and C32 (right panel) EV and SCG2 OE cells before and after IFNγ treatment (10 ng/ml, 48 h). GAPDH was used as a loading control. F The upper panel shows impedance value plotted as the normalized cell index of IFNγ-treated and untreated WM266-4 EV and SCG2 OE cells over time. The lower panel shows the normalized cell index of IFNγ-treated and untreated C32 EV and SCG2 OE cells over time. Graphs show comparisons of the normalized cell index between EV and SCG2 OE, EV and EV treated with IFNγ, EV and SCG2 OE both treated with IFNγ, as well as SCG2 OE and SCG2 OE treated with IFNγ. An increase of the normalized cell index represents cell proliferation and a decrease represents the killing of melanoma cells through T cell-mediated cytotoxicity. EV cells are highlighted in black, IFNγ-treated EV cells are highlighted in grey, SCG2 OE cells are highlighted in red, and IFNγ-treated SCG2 OE cells are highlighted in blue. Cells were treated with10 ng/ml IFNγ for 48 h. Data represent mean ± s.e.m. of three independent experiments (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001; “ns” refers to ≥ 0.05