1. Fig. 4 Lentiviral transgene expression penetrates tissue and provides durable effects in vivo.
Lentiviral transgene expression penetrates tissue and provides durable effects in vivo. (A) Immunohistochemical staining for SOD2 protein expression in SIEA of flaps infected with LVSOD2 [108 transducing units (TUs)] compared with sham [phosphate-buffered saline (PBS)] controls at 180 days after RT. Representative images of n = 3 animals per group are shown. (B) Biochemical assessment of SOD2 activity in flaps infected with LVSOD2 and controls (mean ± SEM) (n = 3 per group). (C) ELISA for CTGF expression in flaps infected with LVSOD2 or LVSOD2 + LVshCTGF (n = 3 per group) compared to controls (mean ± SEM). (D) RT-qPCR for Sod2 gene expression and biochemical SOD2 activity (mean ± SEM) in flap pedicles (artery and vein only) infected with LVSOD2 and controls (n = 3 per group). (E) Immunofluorescent (IF) staining for green fluorescent protein (GFP) expression at 6 months in flaps infected with LVeGFP (108 TUs) and PBS-sham controls (PBS), depicting macrovascular (SIEA and superficial inferior epigastric artery expression; L, lumen; scale bars, 50 μm) and microvascular expression (CD31 colocalization with GFP; scale bars, 25 μm; images representative of three animals per group). Nuclei counterstained with 4′,6-diamidino-2-phenylindole (DAPI). (F) IF staining for extravascular GFP expression (FABP4 colocalization with GFP; scale bar, 50 μm) in flaps infected with LVeGFP (108 TUs; images representative of three animals per group). Nuclei counterstained with DAPI. (G) qPCR for viral gag gene expression in flap tissues infected with LVeGFP at 6 months after infection (n = 3 animals per group). *P < 0.05, **P < 0.01.
Aadil A. Khan et al., Sci Transl Med 2018;10:eaar2041
Published by AAAS