Fig. 6 Combination therapy with LVSOD2 and LVshCTGF preserves flap volume and reduces fibrosis after RT. Combination therapy with LVSOD2 and LVshCTGF preserves.

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Fig. 6 Combination therapy with LVSOD2 and LVshCTGF preserves flap volume and reduces fibrosis after RT. Combination therapy with LVSOD2 and LVshCTGF preserves flap volume and reduces fibrosis after RT. (A) T2-weighted in vivo MRI of irradiated flaps (red arrows) at 180 days after RT (images representative of n = 3 animals per group). (B) MRI-derived volumetric analysis (mean ± SEM) comparing volume changes between groups (n = 3 animals per group). (C) Relative transverse relaxation rate (R2*) differences between groups at 180 days after RT (mean ± SEM) (n = 3 animals per group). (D) Western blot for SOD2 in irradiated and control flap tissues taken from different animals at 180 days after RT with 50 Gy/3 fractions (β-actin loading control). (E) Masson’s trichrome staining for collagen deposition in flap tissues taken at 180 days after RT from each therapeutic group. Images are representative of the larger cohort (n = 6), and whole sections are presented inset (top left). Scale bars, 100 μm. (F) Quantification of Masson’s trichrome staining shown in (E) expressed as total percentage of section exhibiting fibrotic change (mean ± SEM) and correlative RT-qPCR for Col1a2 gene expression in flap tissues 180 days after RT. (G) Multiplexed IF staining for GFP and red fluorescent protein (RFP) expression 180 days after RT in flaps infected with LVSOD2-RFP and LVshCTGF-GFP (both at 108 TUs) or negative controls (PBS) (scale bars, 100 μm). GFP and RFP colocalization is observed within vessel walls (L, lumen). Nuclei are counterstained with DAPI. Cells demonstrating either GFP expression alone (white arrows) or RFP expression alone (red arrows) are also highlighted (images representative of n = 6 animals per group). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Aadil A. Khan et al., Sci Transl Med 2018;10:eaar2041 Published by AAAS