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. 2021 Oct 14;12:6013. doi: 10.1038/s41467-021-26293-w

Fig. 3. NF-YA expression is regulated during satellite cells activation and differentiation.

Fig. 3

a NF-YA immunofluorescence in Pax7+ SCs on freshly isolated EDL myofibers (quiescent, Q), MyoD-activated SCs after 1 day in suspension culture (proliferating, P), MyoG+ differentiating (D) and Pax7+ self-renewing (SR) SCs after 3 days in suspension culture. Nuclei were identified by DAPI staining. n = 3 mice, scale bar: 50 µm. b mRNA levels of NF-YAl and NF-YAs transcripts in isolated SCs measured by RT-qPCR. Data represent mean ± s.d. (two-tailed unpaired t-test: p = 0.0033, *p < 0.05, n = 4 mice). c Protein expression analysis by Western blot of NF-YA and myogenic markers (Pax7, MyoD, MyoG and MyHC) in SC-enriched cultures maintained in proliferating (GM) or differentiating conditions for 1, 3 or 5 days (DM1, DM3, DM5). Tubulin was used as loading control. n = 3 experiments. d Relative mRNA levels of NF-YAl (p = 0.0001, n = 4 mice), Pax7 (p < 0.0001, n = 4 mice) and Myogenin (p = 0.0212, n = 3 mice) in proliferating (GM) and differentiating (DM3) SCs cultured in vitro. Transcript levels in GM condition have been arbitrarily set at 1. Data represent mean ± s.d. (two-tailed unpaired t-test: *p < 0.05, ***p < 0.001, ****p < 0.0001).