Fig. 7: The higher LamR expression in neonatal mice enhances the efficacy of AAV gene therapy in heart.
From: Gene therapy prevents onset of mitochondrial cardiomyopathy in neonatal mice with Ndufs6 deficiency
A The relative expression of hNdufs6 mRNA in cardiac tissue of Ndufs6gt/gt, AAV-Neonatal and AAV-Adult groups at 3 months post AAV gene therapy (n = 4). B Relative protein expression of NDUFS6 in cardiac tissue was detected in Ndufs6gt/gt, AAV-Neonatal and AAV-Adult groups by Western blot. The quantification results were calculated following two steps by Image J: 1) gray scale for each sample was normalized to GAPDH (WT-1 = WT gray scale/GAPDH gray scale; Ndufs6gt/gt-1 = Ndufs6gt/gt gray scale/GAPDH gray scale; AAV-Neonatal-1 = AAV-Neonatal gray scale/GAPDH gray scale; AAV-Adult-1 = AAV-Adult gray scale/GAPDH gray scale); 2) then Ndufts6gt/gt-1, AAV-Neonatal-1 and AAV-Adult-1 are expressed as percentage of WT levels. C Representative images and quantification IHC showed Ndufs6 protein expression in the cytoplasm of heart tissues in Ndufs6gt/gt, AAV-Neonatal and AAV-Adult mice (n = 4). Scale bar=50μm. ***p = 0.0004. D LamR mRNA expression in AC16 cell line after siRNA treatment at different concentration. (n = 3). E LamR protein expression in AC16 cell line after siRNA treatment at different concentration. (n = 3). F Representative images of AC16 cells and AC16-LamR-KD cells 48 h post AAV9-CAG-eGFP infection. Scale bar = 20 um. (n = 3). G LamR mRNA relative expression level in PND0 and 3-month-old mice. (n = 4) H LamR protein expression and quantitative analysis result on PND0 and 3-Month-old mice (n = 4). ***p = 0.0001.
