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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: Stem Cell Res. 2017 Jul 12;23:87–94. doi: 10.1016/j.scr.2017.07.013

Figure 2. PAX7 expression and expansion following multiple MC transfections.

Figure 2

(A) Schematic diagram of differentiation protocol outlining multiple MC transfections (i = CHIR99021; ii–iv = single cell seeding, MC transfections, medium containing FGF2).

(B–C) Graph shows expression levels of GFP measured by FACS in cells that had been subjected to one (x1) or multiple MC transfections, x2, or x3. Cells were analyzed 2 days after each transfection. Non-transfected cells were used as a negative control. Values represent mean ± SEM (n=3 biological replicates) (B). Western blot analysis for PAX7 expression in respective samples (C). ACT was used as housekeeping protein. Lane 1 = D5 non-transfected cells; Lane 2 = D5 MC-transfected cells (x1); Lane 3 = D8 non-transfected cells; Lane 4 = D8 MC-transfected cells (x1); Lane 5 = D8 MC-transfected cells (x2); Lane 6 = D11 non-transfected cells; Lane 7 = D11 MC-transfected cells (x1); Lane 8 = D11 MC-transfected cells (x2); Lane 9 = D11 MC-transfected cells (x3) and Lane 10 = PAX7-induced myogenic progenitors. (n=2 biological replicates).

(D) Growth curve of cells that had been transfected with MC every 3 days, side-by-side with non-transfected cells (negative control) and PAX7-induced myogenic progenitors (positive control). Cell counts were recorded every 4 days, beginning at day 4 (D4). Values represent mean ± SEM. (n=3 biological replicates). *p < 0.05; **p < 0.01; ****p < 0.0001.