Fig. 4

PAS1 suppresses PH20. a The top 10 genes from co-expression analysis were showed. b An empty or PAS1-overexpressing vector was transfected into different breast cancer cells, followed by Western blot. c Control or PAS1 ASOs were transfected into MCF7 cells, followed by Western blot. d Lysates were prepared from MDA-MB-231-control/DNMT1-shRNAs cells for Western blot analysis. e HA-SUV39H1 was transfected into HEK293A cells. RIP assay was performed using anti-HA antibodies, followed by qPCR to detect PAS1 and GAPDH (negative control). f Vectors expressing HA-SUV39H1 and Flag-vigilin were transfected into HEK293A cells, followed by Co-IP assay using anti-HA antibodies. g Biotin-labeled PAS1 was incubated with lysates from MCF7 cells for RNA pull down analysis. h SUV39H1 siRNA was transfected into MDA-MB-231 cells, followed by Western blot. i SUV39H1 was transfected into HEK293A cells. Lysates were extracted for ChIP assays using anti-H3K9me2 and anti-H3K9me3, followed by qPCR to quantify ChIP-enriched DNAs using specific primer for PH20 promoter region. j Lysates were extracted from stable Hs578T-control/PAS1 cells for ChIP assays