Fig. 4
From: MicroRNA regulation of the proliferation and apoptosis of Leydig cells in diabetes
Effect of glucose concentration on miRNAs and apoptosis. Expression of miR-504 (A) and miR-935 (B) in R2C cells at 24 h after culturing in a glucose concentration gradient (basal glucose for R2C cell was 5 mM and stimulated concentrations were 15 mM and 30 mM). Data were normalised to U6 RNA, used as an internal control. Expression of MEK5 (C) and MEF2C (D) determined using RT-qPCR analysis. β-actin was used as an internal control. Representative immunoblotting (E) and cumulative quantification of the protein levels of MEK5 (F) and MEF2C (G) in R2C cells. Media were collected and assayed for concentration of testosterone using ELISA (H). Detection of apoptotic cells using FACS analysis with FITC-labelled annexin V and PI staining (I). Bar graphs represent the percentage of apoptotic cells in each group (J). *p < 0.05, **p < 0.01, ***p < 0.001. n = 3