Research Paper Volume 15, Issue 10 pp 4429—4443

Metformin inhibits ovarian granular cell pyroptosis through the miR-670-3p/NOX2/ROS pathway


Figure 5. Identification of CYBB as a direct target of miR-670-3p. (A) Schematic diagram showing the predicted miR-670-3p binding site in the human CYBB 3’UTR and corresponding mutation sites. (B) Calculation of the minimum free energy hybridization of miR-670-3p and CYBB 3 UTR using RNAhybrid tools; (C) The luciferase reporter plasmids (CYBB-WT and CYBB-Mut) were cotransfected into 293T cells with miR-670-3p mimic or its negative control. Forty-eight hours later, the luciferase activity was detected; (DF) KGN cells were transfected with miR-670-3p mimic/inhibitor or their negative controls for 48 h; (D) RT-PCR analysis of miR-670-3p expression; (E, F) The mRNA and protein levels of NOX2 were determined by RT-PCR and Western blot, respectively. Data are represented as the means ± SD from three independent experiments. ****P < 0.0001, ns. not significant.