FIGURE 3.

Transient heat shock at temperature of 38ºC accelerated myogenic differentiation. (A) Cells were infected twice with SeV‐Myod1 at MOI 64 and then incubated at 37ºC, 38ºC, 39ºC or 40ºC from day 3 to day 8, as shown in the schema in Figure 2A. Myogenic differentiation efficiency was analysed by immunostaining for myosin heavy chain (MHC, green). Nuclei were stained with DAPI (white) to count the total cell number. Scale bar = 100 μm. (B) MHC positivity (%) against total DAPI was quantified and compared among the different temperature conditions. (Independent experiments n = 3, mean ± SD, ***p < 0.001, NS: not significant). (C) MHC‐positive cell area per MHC‐positive cell was quantified and compared among the different temperature conditions. (Independent experiments n = 3, mean ±SD, *p < 0.05, ** p < 0.01, ***p < 0.001). (D) MHC expression levels were analysed and compared among the different temperature conditions. (Independent experiments n = 2, mean ± SD)