Fig. 6
MRVI1-AS1 is induced by hypoxia in a HIF-1-depedent manner in HCC cells. A Hep3B and MHCC-97H cells were exposed to 20% or 1% O2 for 24 h, followed by RT-qPCR (mean ± SD; n = 3). ***P < 0.001, Student’s t test. B JASPAR database was applied to analyze whether there existed potential hypoxia-response element (HRE) in the promoter region of MRVI1-AS1 gene. C Hep3B and MHCC-97H subclones expressing a non-targeting control (NTC) shRNA or a shRNA targeting HIF-1a (sh1a) were exposed to 20% or 1% O2 for 8 h and western blot was performed. D, E Hep3B and MHCC-97H subclones were exposed to 20% or 1% O2 for 24 h, followed by RT-qPCR analysis for MRVI1-AS1 (mean ± SD; n = 3). ***P < 0.001 vs. NTC at 20% O2; ###P < 0.001 vs. NTC at 1% O2 (two-way ANOVA). F–I Hep3B and MHCC-97H cells were exposed to 20% or 1% O2 for 16 h, and ChIP assays were performed by using antibody against HIF-1α or IgG. Primers encompassing HIF binding sites located 0.3 kb 5’, 0.7 kb 5’ to the MRVI1-AS1 transcription start site (TSS) were used for qPCR. Results were normalized to the 848 first lane (mean ± SD; n = 3). ***P < 0.001, two-way ANOVA