Fig. 2.

LncMX1–215 negatively regulates IFNα-induced PD-L1 and galectin-9 expression. a lncMX1–215 expression was detected in HN4 and Cal27 cells via PCR after transfection with expression plasmids for 24 h. b, c The protein and mRNA of PD-L1 and galectin-9 expression levels were detected after transfection with lncMX1–215 for 24 h followed by 200 ng/ml IFNα stimulation for 24 h. d The lncMX1–215 knockdown efficiency after transfection with ASO for 24 h was analyzed. e, f After ASO transfection for 24 h followed by 200 ng/ml IFNα stimulation for 24 h, PD-L1 and galectin-9 expression levels were detected using western blotting and qRT-PCR. g Surface PD-L1 expression was examined using flow cytometry after transfection with lncMX1–215 followed by 200 ng/ml IFNα stimulation for 24 h. h The galectin-9 concentration in medium supernatant was measured via ELISA after transfection for 24 h followed by 200 ng/ml IFNα stimulation for 24 h. i Galectin-9 knockdown efficiency was detected via western blotting after transfection with siRNA for 48 h. j Transfected cells were seeded in a 96-well plate and incubated with NK cells for 4 h at various effector/target (E:T) cell ratios as indicated. The specific lysis rate was measured using an LDH kit. k Relative CD274 mRNA and lncMX1–215 expression were detected using qRT-PCR after 200 ng/ml IFNα stimulation for indicated time. *: P < 0.05; **: P < 0.01