Lentivector-mediated SMN replacement in a mouse model of spinal muscular atrophy
J. Clin. Invest. Mimoun Azzouz, et al. 114:1726 doi:10.1172/JCI22922 [Go to this article.]

Figure 2
Lentivector-mediated restoration of SMN protein expression in vitro. (A) Schematic representation of Lentivector encoding for human SMN gene. CMV, cytomegalovirus; cPPT, central polypurine tract; ΔΔ env, double-deleted envelope leaving only rev response element; WPRE, Woodchuck hepatitis virus posttranscriptional regulation element; SIN, self-inactivating; LTR, long-terminal repeat. (B) Lentivector-mediated expression of SMN in fibroblasts from type I SMA patients. Lentivector-SMN restores SMN expression in gems (arrows). No restoration of gems was observed in fibroblasts incubated with lentivector-LacZ (C). Expression of SMN in human fibroblasts (green) (D) colocalizes with the red immunofluorescence of gemin2 (E), producing yellow staining (F). Arrow indicates colocalization of SMN with gemin2 in gems. Lentivector-LacZ–treated fibroblasts stained with SMN Abs (G) and gemin2 (H). (I) Merged image from G and H. (J) Gem counts in lentivector-SMN–treated and control fibroblast cells. (K) Immunoblot confirming lentivector-mediated SMN expression in human fibroblasts using Abs against SMN. Scale bars: 50 μm (B and C), 100 μm (D–F), 200 μm (G–I).