Fig. 5

SNHG12 regulated HIF1α via endogenous competition with miR-199a-5p. a Alignment between miR-199a-5p seed region and SNHG12 transcript using Starbase online tool. b Luciferase reporter assay was performed to validate the regulatory effect of miR-199a-5p on SNHG12 transcript. Either wild-type or putative binding site mutant SNHG12 was fused to luciferase, which was co-transfected with miR-199a-5p into ACHN and Caki-1 cells. The relative luciferase activities were measured with the Bright-Glo Luciferase Assay System **p < 0.01, ns, no significance. c Correlation between miR-199a-5p and SNHG12 transcripts in renal cancer samples (n = 20). d Alignment between miR-199a-5p and putative target sites in HIF1α 3′UTR by microRNA online tool. e miR-199a-5p negatively modulated HIF1α expression, which was antagonized by SNHG12. Exogenous scramble sequence, miR-199a-5p, SNHG12 or anti-miR-199a-5p were transfected into Caki-1 (left) and ACHN (right) cells in combination as indicated, the relative expression of HIF1α was measured by real-time PCR in indicated cells. f The HIF1α protein was quantified by immunoblotting. β-Actin served as loading control. ***p < 0.001, ****p < 0.0001. g Correlation between miR-199a-5p and HIF1α (upper pane, r = − 0.626, p < 0.01), SNHG12 and HIF1α (lower pane, r = 0.8355, p < 0.0001) in renal cancer samples (n = 20). SNHG12-miR-199a-5p-HIF1α axis was analyzed by q-PCR (h) and western blotting (i) in xenograft tumor. *p < 0.05, **p < 0.01, ***p < 0.001. j Subcellular localization of SNHG12 was analyzed RNA hybridization with the specific Stellaris RNA FISH probes followed by confocal microscope imaging. SNHG12 was detected in red channel, while cytoplasmic GAPDH transcript was detected in green channel. The nuclei were counter-stained with DAPI