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. 2018 Oct 1;13(10):e0204860. doi: 10.1371/journal.pone.0204860

Fig 2. Overexpression of mmu-miR-155 upregulated differentiation associated genes in C2C12 cells.

Fig 2

(A) qPCR analysis for undifferentiated markers (Pax7 and Myf5) and differentiated markers (Mrf4, MyoD, and Myogenin) in mmu-miR-155 overexpressing C2C12 cells at 48 h of transfection (N = 3). Asterisks indicate significant differences at P < 0.05 between control transfected with EGFP control plasmid. (B) WB analysis of the differentiated markers MyoD and Myogenin in mmu-miR-155 overexpressing C2C12 cells at 48 h of transfection. (C) effect of the cumate-induced mmu-miR-155 expression on the marker gene expression in C2C12 cells during differentiation. QM is abbreviation of cumate treatment (N = 3). Asterisks indicate significant differences at P < 0.05. (D) WB analysis for differentiated markers MyoD and Myogenin at day 0 and day 9 of differentiation. (E) immunofluorescence for wildtype (WT) C2C12 cells, C2C12 cells containing the cumate-inducible mmu-miR-155 system (QM-mmu-miR-155) with and without cumate at day 12 after differentiation induction. Scale bars = 100 μm. (F) Myotube formation efficiency at day 12 after differentiation induction with or without cumate. Y-axis of the right figure shows relative differentiation index value to differentiated non-transgenic C2C12 cells (differentiation index value of undifferentiated cells = 0, N = 6). Asterisk means significant difference at P < 0.05.