Figure 6. Restoration of SGCG expression as Mini-Gamma protein.

Reprogrammed fibroblasts were treated for 3 days with either a nontargeting control vivo-PMO (nontarget) or a mutation-specific vivo-PMO exon skipping cocktail (Mini-Gamma). (A) Representative IFM images of reprogrammed 521ΔT cells that were treated as indicated. Treatment with reading frame–correcting vivo-PMOs resulted in the expression of the Mini-Gamma protein (green). Myotubes were labeled with α-actinin (red), and nuclei were labeled with Hoechst 3342 (blue). (B) Image analysis demonstrated a significant increase in Mini-Gamma fluorescence after treatment with vivo-PMOs that corrected the transcript reading frame (n = 6) as compared with nontargeting controls (n = 5). (C and D) Reprogrammed cells harboring a deletion of SGCG exons 5 and 6 (ex5/6del) were treated as indicated. IFM image analysis demonstrated a significant increase in Mini-Gamma fluorescence after treatment with vivo-PMOs that corrected the reading transcript reading frame (n = 4) as compared with nontargeting controls (n = 3). (E and F) Reprogrammed cells harboring a deletion of SGCG exons 6 (ex6del) were treated as indicated. IFM image analysis demonstrated a significant increase in Mini-Gamma fluorescence after treatment with vivo-PMOs that corrected the transcript reading frame (n = 5) as compared with nontargeting controls (n = 3). Data represent the mean γ-sarcoglycan fluorescence per α-actinin–positive area normalized to the mean of the untreated group. A minimum of 3 independent fields were analyzed for each sample. *P < 0.05 as determined by 2-tailed Student’s t test. Data represent the mean ± SEM. Scale bars: 50 μM.