Fig. 5

SPRY4-IT1 functions as a scaffold for PRC2/LSD1/DNMT1 to regulate KLF2 and LATS2 expression in CCA cells. a RT-qPCR detection of the percentage of SPRY4-IT1, GAPDH and U1 in the cytoplasm and nuclear fractions of HuCCT1 and RBE cells. GAPDH and U1 were used as cytoplasmic and nuclear localization markers, respectively. b SPRY4-IT1 levels in immunoprecipitates were determined by RT-qPCR. SPRY4-IT1 expression levels are presented as fold enrichment values relative to IgG immunoprecipitates. c RT-qPCR was performed to confirm the selected gene expression in SPRY4-IT1 knockdown cells. d Western blot assays were performed to confirm the selected gene expression in SPRY4-IT1 knockdown cells. e KLF2 and LATS2 expression levels were examined in cells transfected with EZH2, LSD1 or DNMT1 siRNAs. f ChIP-qPCR analysis of EZH2, H3K27me3, LSD1, H3K4me2 and DNMT1 occupancy on the KLF2 and LATS2 promoter region in HuCCT1 and RBE cells after transfected with si-SPRY4-IT1–1 or si-NC. *P < 0.05, **P < 0.01