Fig. 6: Nocodazole treatment causes accumulation of ribosomes and nascent protein around myonuclei.

A Ribosomes were detected in isolated myofibers using HCR FISH for 28S ribosomal RNA. Representative images of IF/FISH co-labeling for tubulin (green) and telethonin (blue) proteins with rRNA (red) in myofibers. Scale bars: 5 µm. B Active translation was detected in isolated myofibers using in situ puromycylation. Representative images of puromycin detection (red) with tubulin (green) and telethonin (blue) IF in myofibers. Scale bars: 5 µm. C Live myofibers were isolated and cultured with nocodazole or DMSO (control) for 18 or 36 hr. At each time point, rRNA FISH was performed. D Representative images of rRNA FISH signal in myofibers from C. Scale bars: 5 µm. E Perinuclear to cytoplasmic relative rRNA signal density (pixel intensity per µm³) in myofibers from C. Mean ± 95% CI. n = 5 myofibers per condition. *p < 0.05, Two-sided Mann–Whitney U test. F Live myofibers were isolated and cultured with nocodazole or DMSO (control) for 18 or 36 h. At each time point, in situ puromycylation was performed as described in B. G Representative images of puromycin IF in myofibers from F. Scale bars: 5 µm. H Perinuclear to cytoplasmic relative puromycin signal density in myofibers from F. Mean ± 95% CI. n = 5 myofibers per condition. *p < 0.05, Two-sided Mann–Whitney U test.