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Fig. 5 | BMC Biology

Fig. 5

From: TIS7 induces transcriptional cascade of methylosome components required for muscle differentiation

Fig. 5

MSC differentiation is regulated by MyoD, ICln, and TIS7. a TIS7 WT and KO MSCs were plated at the same cell density. Proliferating TIS7 KO MSCs were transfected with indicated plasmids and 6 h later the differentiation was induced. After 5 d of differentiation, cells were fixed, stained with anti-myosin heavy chain antibody MF 20, and the nuclei were visualized using Hoechst. Scale bar = 100 μm. b Percentage of MSC fusion was calculated as the number of nuclei localized in multinucleated (n > 2) myotubes defined by MF20 staining compared to the total number of nuclei. Vertical bars denote the mean ± SD and were calculated from independent biological replicates (n = 3). The indicated number of nuclei was evaluated. Resulting data were analyzed via two-tailed type 2 Student’s t-test, *** P < 0.001. c Quantitative analysis of cells with the indicated nuclei numbers per cell defined by myosin staining. This analysis showed a significant decrease in the number of multinucleated cells and a completely missing category of cells with more than 15 nuclei in TIS7 KO MSCs when compared to TIS7 WT MSCs. Ectopic expression of TIS7, MyoD, or ICln significantly increased the number of multinucleated cells, mainly those with more than 15 nuclei in TIS7 KO MSCs when compared to TIS7 WT MSCs

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