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. 2025 Aug 12;16:1625944. doi: 10.3389/fendo.2025.1625944

Figure 4.

Graphs and images show the effects of miR-204-5p mimics and IGFBP5 on gene expression and cell activity. Panels A and C display bar graphs of relative expression levels of miR-204-5p, IGFBP5, and several mRNAs, showing significant differences marked by asterisks. Panel B illustrates IGFBP5 wild-type and mutant sequences, with a bar graph indicating relative luciferase activity changes. Panel D presents protein expression analyzed through Western blot. Panels E and F include images of cell assays, with corresponding bar graphs showing differences in alkaline phosphatase activity and absorbance at 562 nanometers, highlighting statistically significant changes.

miR-204-5p suppressed the osteogenic differentiation of T2DM-BMSCs by sponging IGFBP5. (A) T2DM-BMSCs were transfected with miR-204-5p mimics or co-transfected with an IGFBP5 overexpression vector, and qPCR was performed to detect the expression levels of miR-204-5p and IGFBP5. (B) A dual-luciferase reporter assay was performed to detect the binding capacity of miR-204-5p and IGFBP5. (C–F) miR-204-5p (and IGFBP5)-overexpressed T2DM-BMSCs were induced for 7 days; osteogenic genes (RUNX2, COL1A1, and BGLAP) were detected using qPCR (C) and western blotting (D); ALP activity was detected using staining and activity quantification (E); and mineralization levels were detected using ARS staining after 14 days of induction (F). BMSCs, bone marrow-derived mesenchymal stem cells; T2DM, type 2 diabetes mellitus. *, P<0.05.