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. 2018 Feb 8;10(3):970–983. doi: 10.1016/j.stemcr.2018.01.007

Figure 6.

Figure 6

p53 Knockdown Rescues Reduced Differentiation and Increased Reserve Cell Generation in Myoblasts Treated with Nutlin-3

(A) Differentiation of C2C12 myoblasts is reduced by Nutlin-3 treatment and rescued by p53 knockdown. C2C12 myoblasts were transfected with either control or p53 siRNA, then maintained for 3 days in low serum supplemented with either DMSO or 20 μM Nutlin-3 prior to fixation and immunostaining to detect myosin heavy chain (MYHC, green) and DNA (DAPI, blue).

(B) Reserve cell (PAX7+/KI67–) generation is increased by Nutlin-3 treatment and rescued by p53 knockdown. C2C12 myoblasts were transfected and cultured as in (A) prior to fixation and immunostaining to detect PAX7 (green), KI67 (red), and DNA (DAPI, blue). Arrows point to PAX7+/KI67+ cells. All PAX7+ cells not labeled by an arrow are KI67–. All KI67+ cells that are not labeled by an arrow are PAX7–.

(C) Quantification of (B) where the average percentage of PAX7+/KI67– cells (over total DAPI+ cells) is calculated across 10–15 technical replicates for 3 independent experiments (N = 35) and plotted. Error bars are SEM. ∗∗p < 0.01.