Fig. 6

Effect of miR-29c-3p in ECM composition, osteogenic differentiation, cell proliferation and apoptosis. a COL1 staining and quantification at day 7 of differentiation. (COL1: red; N = 6; scale: 100 μm) b Expression of ECM components (COL1A1, COL1A2 and COL3A1) by qPCR after 3 days in osteogenic-inducing conditions (N = 6). c Correlation between miR-29c-3p expression level and collagen expression on bone samples. d Expression levels of osteogenic markers at day 3 after transfection and incubation in osteogenic-inducing conditions (N = 6). e ALP and f Alizarin staining (N = 6) and Alizarin solubilization quantification (N = 6), after transfection and culture for 7 and 14 days, respectively, in osteogenic-inducing conditions. g Representative fluorescence profile (resorufin) measured every 24 h for 7 days in transfected MSCs and quantification of the percentage of cells in proliferation (Ki-67⁺/DAPI⁺) after 3 days in culture (N = 6). h Quantification of viable (Annexin V⁻ PI⁻), early apoptotic (Annexin V⁺ PI⁻), or late apoptosis and dead MSCs (Annexin V⁻ PI⁺ and Annexin V⁺ PI⁺) (N = 6). (Mimics-NC: MSC transfected with scrambled miRNA mimics negative control; Mimics: miR-29c-3p transfected MSC; Inhibitor-NC: MSC transfected with scrambled miRNA inhibitor negative control; Inhibitor: miR-29c-3p-inhibitor transfected MSC).