Fig. 4

MIAT regulated CDKN1B via endogenous competition with miR-150. a Alignment between miR-150-5p seed region and MIAT transcript (wild-type and mutant) using Starbase online tool. b, c Luciferase reporter assay was performed to validate the regulatory effect of miR-150 on MIAT transcript. Either wild-type or putative binding site-mutant MIAT was fused to luciferase, which was co-transfected with miR-150 into HeLa and HT-3 cells. The relative luciferase activities were measured with the Bright-Glo Luciferase Assay System. n.s: no significance, *p < 0.05, **p < 0.01; d Correlation between miR-150 and MIAT transcripts in cervical cancer samples (n = 21). e Alignment between miR-150 and putative target sites in CDKN1B 3′UTR by microRNA online tool. f miR-150 negatively modulated CDKN1B expression, which was antagonized by MIAT. Exogenous scramble sequence, miR-150, MIAT or anti-miR-150 were transfected into HeLa (left) and HT-3 (right) cells in combination as indicated, the relative expression of CDKN1B was measured by real-time PCR in indicated cells. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. g The CDKN1B protein was quantified by immunoblotting. β-actin served as loading control. h, i The MIAT-miR-150-CDKN1B axis was analyzed by q-PCR (h) and western blotting (i) in xenograft tumor. *p < 0.05, **p < 0.01, ****p < 0.0001. j The relative expression of both miR-150 and CDKN1B were analyzed in paired human cervical tumor samples by real-time PCR