Fig. 9

Silencing of SAT1 alleviates FMP-Induced Pulmonary Fibrosis in vivo. Embryonic day 2 (ED2) chicken embryos were micro-injected with SAT1-silencing lentivirus, and then 24 h-old-chickens were exposed to 0.6 mg/kg FMP via aerosol inhalation. SAT1 expression in chicken lung tissues were assessed with qRT-PCR. Extent of pulmonary fibrosis were determined with H&E, Masson and Sirius red staining. Protein expression levels of ALOX15, SAT1, GPX4, HO1 and TGF-β were assessed with western blotting. Data were expressed as mean ± SD, N = 5 per group; * indicates significant difference from the blank control group (p < 0.05); ** indicates significant difference from the blank control group (p < 0.01); # indicates significant difference from 0.6 mg/kg FMP exposure group (p < 0.05); ## indicates significant difference from 0.6 mg/kg FMP exposure group (p < 0.01). (A) Schematic of animal treatments and qRT-PCR confirmation of SAT1 silencing efficacy (N = 4). (B) Representative pictures of chicken lung tissue sections from animals exposed to 0.6 mg/kg FMP, with or without SAT1 silencing, Yellow arrows represent positive areas. (C) Quantification of Ashcroft score and fibrotic lesion ratio for the chicken lung tissue sections from animals exposed to 0.6 mg/kg FMP, with or without SAT1 silencing (N = 5). (D) Representative western blotting images for ALOX15, SAT1, GPX4, HO1 and TGF-β in the lung tissues samples from animals exposed to 0.6 mg/kg FMP, with or without SAT1 silencing, along with semi-quantifications, The quantification of GPX4 bands was performed using N = 5 animals, while the remaining proteins were quantified with N = 4 animals