Fig. 3

MRVI1-AS1 increases SKA1 expression by stabilizing SKA1 mRNA in HCC cells. A Heat map for downstream genes of MRVI1-AS1 in MHCC-97H cells. Among the downstream genes of MRVI1-AS1, the expression fold change of SKA1 is remarkable upon MRVI1-AS1 knockdown. B, C RT-qPCR and western blot were performed to detect mRNA and protein levels of SKA1 in MRVI1-AS1-knockdown subclones of MHCC-97H (mean ± SD; n = 3; ***P < 0.001, two-way ANOVA) and Hep3B cells (mean ± SD; n = 3; ***P < 0.001, Student’s t test). D RT-qPCR was conducted to explore the expression of SKA1 in HCC tissues (n = 72) and adjacent non-tumor (NT) tissues (n = 72). ***P < 0.001, Student’s t test. E TCGA data from UALCAN platform showed that MRVI1-AS1 expression in HCC tissues (n = 371) was significantly higher than that in normal tissues (n = 50). Student’s t test. F Pearson correlation analysis was applied to examine the correlation between MRVI1-AS1 and SKA1 mRNA in HCC tissues. G Data from UALCAN platform showed that HCC patients in high SKA1 expression group had worse prognosis. H Luciferase reporter assay was applied to detect whether MRVI1-AS1 influence the luciferase activity of SKA1 promoter (mean ± SD; n = 3). Student’s t test. I The separation of nuclear and cytosolic fractions assay was applied to determine the subcellular localization of MRVI1-AS1 in HCC cells (mean ± SD; n = 3). J, K HCC cells with MRVI1-AS1 alteration were treated with actinomycin D (Amyjet Scientific, Wuhan, China) to block RNA synthesis, and the degradation of SKA1 mRNA was examined using RT-qPCR assay at different time point (mean ± SD; n = 3). **P < 0.01, **P < 0.001, two-way ANOVA