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. 2024 Oct 28;9:32. doi: 10.1038/s41536-024-00376-w

Fig. 2. No Mest-induced in vitro MSC multipotency.

Fig. 2

A SPRING plot of the integrated scRNAseq fibroblasts, (A) colored by Seurat clusters, (A’) time point, and (A”) Mest expression in green; MSK musculoskeletal; CT connective tissue. B–D Oil Red O staining of (B) pCAG-GFP C3H10T1/2, (C) pCAG-GFP MC3T3, and (D) pCAG-Mest MC3T3 cells at 12 dpi. E Quantification of extracted Oil Red O from 12 dpi stained MC3T3 cells in (C) and (D). F, G Adipogenesis qPCR analysis of (F) Perilipin-1 and (G) PPARγ genes in GFP (gray) and Mest (black) transfected MC3T3 cells at 1, 7, and 12 dpi. H–J Alcian Blue staining of (H) pCAG-GFP C3H10T1/2, (I) pCAG-GFP MC3T3, and (J) pCAG-Mest MC3T3 cells at 21 dpi. K Quantification of Alcian Blue stain extracted from 21 dpi stained MC3T3 cells in (I) and (J). L, M Chondrogenesis qPCR analysis of (L) Col2a1 and (M) Col10a1 genes in GFP and Mest transfected MC3T3 cells at 7, 14, and 21 dpi. Error bars depict standard deviation. Statistical comparisons are shown for those discussed in text; (**) p < 0.01, (***) p < 0.001, (****) p < 0.0001, ns not significant. Scale bars in images (B–D) = 100 µm, scale bars in images (H–J) = 250 µm.