Fig. 6

UPK1A-AS1 interacts with EZH2. a. UPK1A-AS1 cytosolic and nuclear expression levels in SK-Hep-1 and MHCC-97H cells. β-Actin and U6 were used as cytosolic and nuclear markers, respectively. Both NEAT1 and MALAT1 were localized to the nucleus. b-c. RIP assay was performed in MHCC-97H cells and the co-precipitated RNA was subjected to qRT-PCR for UPK1A-AS1(^***P < 0.001). d. RNA pull-down assay was carried out to confirm the association between UPK1A-AS1 and EZH2. e. Effect of UPK1A-AS1 overexpression on EZH2 and H3K27M3 expressions was measured by western blotting. f. Correlation between UPK1A-AS1 and EZH2 was analyzed in HCC samples from TCGA dataset. g. EZH2 expression level in the cytoplasm or nucleus of MHCC-97H cells. β-Actin was used as a cytosol marker, LaminB1 served as a nuclear marker. h. Translocation of EZH2 from the cytoplasm to the nucleus was detected by immunofluorescence assay. i. Immunoprecipitation assay identified the increased interaction between EZH2 and SUZ12 in UPK1A-AS1-overexpressing MHCC-97H cells. β-Actin was used as negative control