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. 2022 Jun 8;13:3180. doi: 10.1038/s41467-022-30778-7

Fig. 2. Loss of miR-1/206/133 in skeletal muscles disturbs neuromuscular synapse formation and causes respiratory failure.

Fig. 2

a Lung floating test of control and miR-1/206/133 tKO reveals deficits in breathing after recovery by caesarean section in tKO animals compared to control littermates [E18.5]. b Representation of percentages of control and miR-1/206/133 tKO in the lung floating test (percent of floating lungs in each group; n = 28 control/11 tKO from 8 litters, control = littermates [E18.5]). c Measurement of average endplate band width of control and miR-1/206/133 tKO diaphragms (n = 3 control/4 tKO, control = littermates, Mann-Whitney-U test, one-tailed, *p = 0.0286, [E18.5]). d Whole-mount bungarotoxin (BTX) staining of diaphragms from control and miR-1/206/133 tKO animals (control = littermates, scale-bar: 600 µm upper row/50 µm lower row [50 µm thick Z-stacks lower row, n = 3 control/4 tKO, [E18.5]). e Whole mount BTX staining of hemi-diaphragms of miR-206/133b sKO (n = 3) and miR-1/133a dKO (n = 2) animals (scale-bar: 600 µm upper row/100 µm lower row [E18.5]). f Z-stacks of a whole mount-stained diaphragms from control (littermate) and miR-1/206/133 tKO mice. Anti-SV2 (synaptic vesicles) and anti-neurofilament antibodies were used to label both nerve terminals and axons (green). Staining of the postsynapse was done using BTX (red). Arrows indicate AChR clusters without a nerve terminal in close proximity (scale-bar: 20 µm, n = 3 control/3 tKO [E18.5]). g, h Quantification of the average AChR cluster number and AChR volume in BTX-stained whole mount diaphragms based on Z-stacks, comparing miR-1/206/133 tKO and control samples (Supplementary Fig. 5b). Data were obtained from 3D rendered confocal Z-stacks (n = 3 control/3 tKO, control = littermates, Mann-Whitney-U test, one-tailed, *p = 0.05, [E18.5]). Source data are provided as a Source Data file. Data are presented as mean values +/− SEM in c and g, h.