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Fig. 5 | Nutrition & Metabolism

Fig. 5

From: Salvianolic acid B activates chondrocytes autophagy and reduces chondrocyte apoptosis in obese mice via the KCNQ1OT1/miR-128-3p/SIRT1 signaling pathways

Fig. 5

KCNQ1OT1 is a ceRNA of miR-128-3p in the regulation of SIRT1 expression. A The binding sites of KCNQ1OT1 and miR-128-3p were predicted by bioinformatics analysis. B Dual luciferase reporter assay was performed to detect the luciferase activity of HEK-293 T cells co-transfected with miR-128-3p mimic and luciferase reporter vectors containing WT- or MUT- KCNQ1OT1 3’UTR. C The enrichment of KCNQ1OT1 and miR-128-3p in IgG or Ago2 immunoprecipitates detected by RIP and RT-qPCR. Anti-IgG served as a negative control, and 30% Input (cells extracts) served as a positive control. D Relative expression levels of miR-128-3p in ATDC5 cells transfected with KCNQ1OT1 shRNA, Ad-KCNQ1OT1 or their negative controls. E Expression of miR-128-3p in cartilage tissues obtained from normal group and OA group was determined. F The correlation analysis between KCNQ1OT1 and miR-128-3p was determined by spearman analysis. G The putative binding sites between miR-128-3p and SIRT1. H and I Dual-luciferase reporter gene assay and RIP analysis were performed to verify the correlation of miR-128-3p and SIRT1. Anti-IgG served as a negative control, and 30% Input (cells extracts) served as a positive control. J and K Expression levels of miR-128-3p and SIRT1 protein were detected in ATDC5 cells transfected with miR-128-3p mimic, miR-128-3p inhibitor and their negative control. Data are expressed as means ± SEM. Student’s t test was used for the comparison between 2 groups in this study. *P < 0.05

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