SUV420H2 silences MET transcription factors via the H4K20me3 repressive mark. (A) qRT-PCR expression analysis of CDH1, EPCAM, and VIM in PANC-1 cells in double-knockdown rescue experiments. Data are normalized to siNTC control; bar graphs indicate mean ± SD. n = 3 biological replicates each averaged from three technical replicates; differences were assessed by Student’s t test compared with siNTC control, unless otherwise indicated. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (B) Mass spectrometry analysis of global K4K20 methylation patterns in PANC-1 cells with control or SUV420H2 knockdown. Black values above the bars represent the relative occurrence compared with the siNTC control for each mark; white values below the bars represent the absolute values from the mass spectrometry assay. (C) Western blot for global H4K20me3 levels in mesenchymal pancreatic cancer cells with siNTC control and SUV420H2 knockdown. H4 is the loading control. (D) ChIP-qPCR results quantifying levels of H4K20me3 on FOXA1, OVOL1, and OVOL2 genes in PANC-1 cells, with control or SUV420H2 knockdown. Gene diagrams include genomic locus information. Three regions per gene were chosen for probing, shown as pink lines and numbers, indicating the location relative to the gene sequence. Each dot represents a value for one biological replicate, averaged from three technical replicates. Black lines indicate means. Differences were assessed by Student’s t test compared with siNTC control for each probe. *, P < 0.05; **, P < 0.01; ns, not significant (P ≥ 0.05).