Supplementary Figure I

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Supplementary Figure I AAV9-CMV- EGFP AAV9-CMV- EGFPmiR122(3x)TS AAV9-CMV-MLC0.26- EGFP AAV9-CMV-MLC0.26- EGFPmiR122(3x)TS heart Supplementary Figure I

Supplementary Figure II AAV9-CMV- EGFP AAV9-CMV- EGFPmiR122(3x)TS AAV9-CMV-MLC0.26- EGFP AAV9-CMV-MLC0.26- EGFPmiR122(3x)TS femoris muscle Quadriceps Supplementary Figure II

Supplementary Figure III AAV9-CMV- MLC0.26-EGFP AAV9-CMV- MLC0.26-EGFP AAV9-CMV- EGFP AAV9-CMV- EGFP AAV9-CMV- EGFPmiR122(3x)TS AAV9-CMV- EGFPmiR122(3x)TS AAV9-CMV-MLC0.26-EGFPmiR122(3x)TS AAV9-CMV-MLC0.26-EGFPmiR122(3x)TS Supplementary Figure III

Supplementary Figure Legends Supplementary Figure I: Regulation of AAV9 vector mediated gene expression by miR122 in the heart does not affect transduction of cardiomyocytes. Representative microphotographs show colocalization of EGFP fluorescence with an actin stain using phalloidin (red). The pattern of EGFP expression in the heart is similar between all groups irrespective of the use of miR122 TS. Bar: 50 µm. Supplementary Figure II: Visualization of EGFP expression in skeletal muscle. Cryosections of quadriceps femoris muscle from adult mice 4 weeks after intravenous injection of 7.5x1011 vg of AAV9-CMV-EGFP, AAV9-CMV-EGFPmiR122(3x)TS, AAV9-CMV-MLC0.26-EGFP, and AAV9-CMV-MLC0.26-EGFPmiR122(3x)TS were compared. Expression was found to be close to the detection limit. No difference in expression was detectable either with or without miR122 TS in combination with the CMV or CMV-MLC0.26 promoter. Bar: 50 µm Supplementary Figure III: Comparison of AAV vector DNA amount in vivo. Vector DNA of AAV9-CMV-EGFP, AAV9-CMV-EGFPmiR122(3x)TS, AAV9-CMV-MLC0.26-EGFP and AAV9-CMV-MLC0.26-EGFPmiR122(3x)TS in heart and liver was quantified by real-time PCR and related to GAPDH DNA. Shown are DCt values of vector DNA vs. GAPDH DNA, the mean and standard error of mean (SEM). The vector DNA levels measured in the heart and the liver were very similar between all groups.