Fig. 6

Knockdown of NEAT1 down-regulates ATGL expression through upregulation of miR-124-3p levels. a Predicted conserved miR-124-3P binding sites of NEAT1 and ATGL. The calculated free energy value of the hybrid is indicated. The generated mutant sites in NEAT1 (or ATGL) 3′ UTR seed regions are indicated. b Real-time PCR analysis showing the effect of sh-NEAT1 on miR-124-3p expression in HCCLM3 and SK-Hep-1 cells. c Dual-luciferase reporter assays revealed that co-transfection of NEAT1-WT with miR-124-3p in SK-Hep-1 cells decreased luciferase activity as compared with co-transfection using NEAT1 muture. d Real-time PCR and western blot analysis showing the effect of up-regulation of miR-124-3p using mimic on ATGL expression in HCCLM3 and SK-Hep-1 cells. e Real-time PCR and Western blot results revealed that inhibition of NEAT1 down-regulated ATGL expression, and this suppression was attenuated in cells with inhibited miR-124-3p. f Dual-luciferase reporter assays revealed that co-transfection of ATGL-WT with miR-124-3p in SK-Hep-1 cells decreased luciferase activity as compared to co-transfection using ATGL muture. g Correlation analyses revealed that expression levels of NEAT1 (or ATGL) were inversely associated with those of miR-124-3p in 40 clinical HCC samples. Pearson correlation coefficient was used to as a measure of association. Data are expressed as mean ± SD of three independent experiments. Statistical significance was concluded at *P < 0.05, **P < 0.01, ***P < 0.001; NS represents no statistical significance