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. 2021 Aug 19;12:5043. doi: 10.1038/s41467-021-25170-w

Fig. 9. GNAI2 mediates DHX36 function in SC proliferation.

Fig. 9

a C2C12 cells were transfected with siNC or siGnai2 oligos and the successful deletion of GNAI2 protein was detected by western blot with α-Tubulin as the internal loading control. b SCs were transfected with the above siRNA oligos for EdU assay. The percentage of EdU incorporation was quantified from 10 randomly selected fields/sample. Scale bar = 100 μm; n = 3 independent experiments. per group. c Ctrl SCs were infected with GFP or Gnai2-expressing lentivirus and the Gnai2 overexpression was confirmed by western blot. d EdU assay was conducted with the above cells and the relative EdU incorporation percentage was quantified from ten randomly selected fields per sample, with the value from GFP virus-infected cells set as 1. Representative images are shown. Scale bar = 100 μm; n = 3 mice per group. e iKO SCs were infected with GFP or Gnai2-expressing lentivirus and the expression of GNAI2 protein was confirmed by western blot at 48 h after infection. f EdU labeling was performed as above described and the incorporation percentage was quantified from ten randomly selected fields per sample. Representative images are shown. Scale bar = 100 μm; n = 3 mice per group. g Dhx36-KO myoblast cells were transfected with pcDNA3.1 vector plasmid or pcDNA3.1-mDhx36 overexpression plasmid for 48 h and the expression of Dhx36 and GNAI2 protein was confirmed by western blot. h Ctrl or Dhx36-iKO SCs were transfected with vector or pcDNA3.1-Dhx36 overexpression plasmid for 36 h and EdU was added to the cells 4 h before harvesting. The relative EdU incorporation percentage was quantified from ten randomly selected fields per sample. Scale bar = 100 μm; n = 3 mice per group. i Schematic illustration of the functional mechanism of DHX36 in regulating SC proliferation and muscle regeneration. Data represent mean ± s.d. (b, d, f, h) and Student’s t test (two-tailed unpaired) was used to calculate the statistical significance in b, d, f, h (iKO group); Student’s t test (two-tailed paired) was used to calculate the statistical significance in h (Ctrl group): *P <  0.05; **P < 0.01. Source data are provided as a Source Data file.