Fig. 4: SpCas9 mRNA-mediated reframing efficiently rescues dysferlin protein expression and correct localization in patient MuSC while maintaining their myogenic profile. | Nature Communications

Fig. 4: SpCas9 mRNA-mediated reframing efficiently rescues dysferlin protein expression and correct localization in patient MuSC while maintaining their myogenic profile.

From: Gene-editing in patient and humanized-mice primary muscle stem cells rescues dysferlin expression in dysferlin-deficient muscular dystrophy

Fig. 4

a Western blot analysis of dysferlin protein expression in edited patient MuSC. Unedited patient and control MuSC were used as controls. Vinculin was used as loading control. b The intensity of the dysferlin bands from a was quantified using ImageJ and normalized to Vinculin (n = 3 replicates, mean ± SD). c Dysferlin immunostaining of terminally differentiated control and patient MuSC transfected with SpCas9 mRNA, with or without sgRNA#3. Dysferlin membrane localization is indicated by white arrows. Scale bars: 25 µm. d Edited patient MuSC stained for DES, PAX7 and MYOD. Scale bar: 50 μm. e Fold-change of the percentage of patient cells expressing myogenic and proliferation markers at day 5 after transfection of SpCas9 mRNA, with or without sgRNA (normalized to untransfected cells) (n = 1 independent experiment per patient, mean ± SD). Source data are provided as a Source Data file.

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