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FIGURE 9. miR‐466m‐5p–mediated regulation of MSC osteogenesis. (a) ALP staining of MSCs transfected with miR‐466m‐5p mimics or miR‐NC on days 3, 5, and 7 of osteogenic induction. (b) Alizarin Red S staining of MSCs transfected with miR‐466m‐5p or miR‐NC on days 7 and 14. (c) quantitative assessment of ALP activity in MSCs transfected with miR‐466m‐5p or miR‐NC on days 3, 5, and 7. (d) semi‐quantitative analysis of Alizarin Red S staining in MSCs transfected with miR‐466m‐5p or miR‐NC. (e) mRNA expression levels of osteogenic genes COL1A1, ALP, OCN, OPN, and OSX on days 1, 3, and 7 after transfection. (f) Flow cytometric analysis of apoptosis in MSCs transfected with miR‐466m‐5p or miR‐NC and exposed to different concentrations of LPS (0, 0.5, 1, 2, 4, 8, 16, and 32 µg/mL). (g) Quantitative analysis of apoptotic cells based on flow cytometry. (h,i) Immunofluorescence staining of RUNX2 (h) and COL1A1 (i) in MSCs transfected with miR‐466m‐5p or miR‐NC. Quantitative analysis of RUNX2 (j) and COL1A1 (k) fluorescence intensity. Image published in: Lu Y et al. (2026) Image downloaded from an Open Access article in PubMed Central. © 2026 The Author(s). Endocrinology, Diabetes & Metabolism published by John Wiley & Sons Ltd. Larger Image Printer Friendly View

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