Fig. 1: Activation-induced DHX36 is required for muscle formation.
a Schematic for isolation of quiescent satellite cells (SCT0) after in situ fixation, freshly isolated SCs without prior fixation (SCT8) and SCs cultured for 24, 48, or 72 h. b RNAs from the above cells were subject to RNA-seq and FPKM of Dhx36 mRNA is plotted. c DHX36 protein in the above SCs were examined by western blot. The relative intensity of each band is calculated by normalizing with total proteins stained by Ponceau S. The intensity for SCT8 was set as 1. d Mouse TA muscles were injected with barium chloride (BaCl2) and the muscle homogenates were collected at the designated dpi for qRT-PCR measurement of Dhx36 mRNAs. n = 3 independent experiments; P = 0.0000097 and 0.000015. e Immunofluorescence staining of DHX36 protein in SCT0, SCT8 or SC 24 h. Scale bar: 2 μm. f Breeding strategy to generate Dhx36 cKO mouse. g H&E staining of TA muscle from adult Ctrl or Dhx36 cKO mice. Scale bar = 100 μm (×20 images) or 50 μm (×40 images). h Average fiber size in the above muscles was measured, Ctrl: n = 3 mice; cKO: n = 4 mice; P = 0.003. i Cross-section area (CSA) of each fiber from the above muscles (Ctrl in white and cKO in gray) was quantified and the distribution is shown. From left to right, P = 0.014, 0.027, 0.000013, 0.008, 0.003, 0.026, 0.007. j Grip strength of fore and hindlimb muscles were measured in Ctrl and cKO mice, Ctrl: n = 8 mice; cKO: n = 7 mice; P = 0.023. k Immunohistochemistry staining of eMyHC was performed on sections of TA muscles from Ctrl and cKO mice on 7 dpi, scale bar = 100 μm. l The average fiber size of newly formed fibers was quantified from the above eMyHC+ fibers, Ctrl: n = 3 mice; cKO: n = 4 mice; P = 0.022. m Distribution of CSA of the above fibers (Ctrl in white and cKO in gray). P = 0.011 and 0.033. Data are represented as mean ± s.d., Student’s t test (two-tailed unpaired) was used to calculate the statistical significance (d, h, i, j, l, m): *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.
