Fig. 4

Bioinformatics analysis, dual luciferase assay and RIP experiments confirmed the interaction of ZFPM2-AS1 with miR-139. a Bioinformatics analysis predicted that ZFPM2-AS1 has a negative correlation with miR-139. b PCR detection of the expression levels of miR-139 in wild type or ZFPM2-AS1 silenced Huh7 and HCCLM3 cells. c Luciferase reporter assay to detect the luciferase activity of pmiR-ZFPM2-AS1 in Huh7 and HCCLM3 cells cotransfected with miR-139-mut/wt. d IP-western detection of anti-Ago2 antibody capture in HCCLM3 cell lysates, with IgG as the negative control and Input as the positive control. PCR and qRT-PCR was both used to detect the expression of ZFPM2-AS1 and miR-139 in the immunoprecipitated and the same expression trends were observed. IgG was the negative control and Input was the positive control. Data are expressed as the mean ± standard deviation, ** p < 0.01, *** p < 0.001. (e_left) qPCR validation of ZFPM2-AS1 and miR-139 enrichment versus input after ZFPM2-AS1 RNA pull-down by specific probes (SP) as compared to non-specific probes (NSP) in HCCLM3 cells. (e_right) qPCR validation of miR-139 and ZFPM2-AS1 enrichment versus input after miR-139 RNA pull-down by specific probes (SP) as compared to non-specific probes (NSP) in HCCLM3 cells(**p < 0.01)