Figure 3.

GAS5 bound to miR-382-3p to positively regulate CYP2C8 expression. (a) Top three lncRNAs (SNHG1, GAS5 and NORAD), which could possibly associate with miR-382-3p, were presented using ENCORI. Luciferase reporter assay evaluated the the luciferase activity of SNHG1-WT, GAS5-WT and NORAD-WT reporters after the transfection of NC-mimics or miR-382-3p mimics into HepG2 and Huh-7 cells. (b) FISH assay revealed GAS5 subcellular location in HepG2 and Huh-7 cells. (c) Cytoplasmic and nuclear RNA isolation uncovered the distribution of GAS5 in the cytoplasm or nuclei of HepG2 and Huh-7 cells. (d) lncLocator predicted a high possibility of the cytoplasmic abundance of GAS5. (e) RIP assay illustrated the enrichment of GAS5, miR-382-3p and CYP2C8 with Ago2 protein. (f) Complementary sequences between miR-382-3p and GAS5, together with the mutant site of GAS5 were shown. (g) Luciferase reporter assay showed the decline of GAS5-WT reporter luciferase activity due to the transfection of miR-382-3p mimics. (h) The impact of GAS5 overexpression on miR-382-3p level in LC cells was assessed by qRT-PCR. (i) qRT-PCR and western blot illustrated the effects of altered GAS5 expression on CYP2C8 mRNA and protein levels in LC cells. p < .01 (**), p < .001 (***), N.S.: not significant