Fig. 5

LINC00152 interacted with EZH2, and regulated IL24 expression. a qRT-PCR was performed to detect the relative LINC00152 levels in A549 and SPCA1 cell cytoplasm and nucleus. GAPDH was used as cytoplasm control and U6 was used as nucleus control. b RIP experiments were performed in A549,SPCA1 cells and the corprecipitated RNA was subjected to qRT-PCR for LINC00152. Expression levels of LINC00152 RNA are as fold enrichment in EZH2,LSD1,SUZ12 relative to IgG immunoprecipitates. c RNA pulldown and western blotting assays revealed biotinylated LINC00152 could bind to EZH2 and LSD1. d qRT-PCR was used to analyze the mRNA expression levels of tumor suppressor genes in scrambled and si-LINC00152 2#/3# treated LAD cells. e Western blotting analysis of IL24 protein level in LINC00152knockdown A549 and SPCA1 cells, GAPDH protein was used as an internal control. f IL24 mRNA level in A549 and SPCA1 cells transfected with si-EZH2. g Western blotting were performed to detect the IL24 protein level in A549 and SPCA1 cells. h ChIP-qRT-PCR of EZH2 occupancy and H3K27me3 binding in the IL24 promoter in A549 and SPCA1 cells treated with si-LINC00152(48h) or scrambled siRNA, IgG was used as a negative control. Values are shown as the mean ± s.d in three independent experiments. *P < 0.05, **P < 0.01