REP1 expression from the vector AAV2/5-CAG-CHM. (a) Immunofluorescence studies of REP1 expression (in green), nuclei in blue, and F-actin filaments in red in wild-type fibroblasts. (b) Absence of REP1 expression in CHM1 fibroblasts. (c) REP1 expression (in green) from AAV2/5-CAG-CHM 48 hours posttransduction of CHM1 fibroblasts with 100,000 vector genomes (vg) per cell. Bars = 50 µm. (d) Western blot analysis of fibroblasts 48 hours posttransduction with 100,000 vg per cell of AAV2/5-CAG-CHM showed that REP1 was expressed at a level ~17% of wild-type (WT) expression level. (e) Western blot analysis of RPE 4 weeks posttransduction with 100,000 vg per cell of AAV2/5-CAG-CHM showed that REP1 was expressed at a level ~53% of wild-type level. Note the absence of REP1 expression in nontransduced (NT) CHM1 cells in both d and e. AAV, adeno-associated vector; CAG, chicken β-actin with a CMV enhancer; REP1, Rab escort protein 1; RPE, retinal pigment epithelium.