Fig. 4

LncMX1–215 directly interacts with GCN5. a GCN5 and H3K27ac were detected after transfection with lncMX1–215 for 24 h and treatment with 200 ng/ml IFNα for 24 h. b GCN5 and H3K27ac were detected after transfection with ASO for 48 h. c, d GCN5, PD-L1, galectin-9 and H3K27ac were examined after transfection with lncMX1–215 or ASO for 24 h followed by treatment with 5 μM MS-275 for 24 h. e GCN5, PD-L1, galectin-9 and H3K27ac were detected after ectopic expression of GCN5 for 48 h in HN4 and Cal27 cells. f PD-L1 and galectin-9 were detected after cotransfection with GCN5 and lncMX1–215 for 48 h. g After transfection with lncMX1–215 for 48 h, cell lysates were precipitated with anti-GCN5 or IgG antibody. h GCN5 and H3K27ac expression was analyzed and quantified using immunofluorescence. i The correlation between H3K27ac and GCN5 and lncMX1–215 expression was analyzed in HNSCC tissue microarray. j Overexpression of GCN5-DYKDDDDK fusion plasmid in HN4 cells was verified by western blotting. P1 to P5 were truncations for full length of lncMX1–215 amplified by specific primers. k After transfection of HN4 cells with GCN5-DYKDDDDK fusion plasmid for 48 h, RNA immunoprecipitation was used to detect binding of GCN5 and lncMX1–215. The binding fragments were detected using PCR with primers P1 to P5. l Cell lysates were incubated with biotinylated lncMX1–215 or an unrelated probe, and the eluent and flow-through were analyzed via western blotting with anti-GCN5 antibody. L: lysate load; FT: flow-through; E: eluent. *: P < 0.05; **: P < 0.01