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. 2022 Aug 29;54(8):1277–1289. doi: 10.1038/s12276-022-00831-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — Eight-week-old f/f; Alb-Cre mice were infected by adenovirus expressing GFP or TBG-SMAD7. The mice underwent sham operation (n = 3) or BDL (n = 5–9). A SMAD7 expression in the liver was determined by immunoblotting 3 days after BDL. B Seven days after BDL, the liver was analyzed by H&E and Sirius Red staining and immunofluorescence analysis of αSMA and CK19, p-SMAD2/3, and SMAD7. C Liver fibrosis was quantified by measuring the area stained with Sirius Red. Two to three images of each liver showing Sirius Red staining were obtained. D Each specimen was analyzed by immunoblot analysis of p-SMAD2/3, SMAD2/3, SMAD4, αSMA, cleaved Caspase-3, and GAPDH. E The band intensities of p-SMAD2/3, αSMA, and cleaved Caspase-3 in D were statistically analyzed after normalization to the band intensity of GAPDH. F The mRNA levels of Smad7, Acta2, Col1a1, and Krt19 in each liver specimen were analyzed by RT–qPCR. The mRNA levels were normalized to those of 18S rRNA. TBG, thyroxine-binding globulin. Scale bar = 20 µm. *p < 0.05, **p < 0.01, ***p < 0.001. All data are the mean ± S.E.M.

Fig. 7 — Eight-week-old f/f; Alb-Cre mice were infected by adenovirus expressing GFP or TBG-SMAD7. The mice underwent sham operation (n = 3) or BDL (n = 5–9). A SMAD7 expression in the liver was determined by immunoblotting 3 days after BDL. B Seven days after BDL, the liver was analyzed by H&E and Sirius Red staining and immunofluorescence analysis of αSMA and CK19, p-SMAD2/3, and SMAD7. C Liver fibrosis was quantified by measuring the area stained with Sirius Red. Two to three images of each liver showing Sirius Red staining were obtained. D Each specimen was analyzed by immunoblot analysis of p-SMAD2/3, SMAD2/3, SMAD4, αSMA, cleaved Caspase-3, and GAPDH. E The band intensities of p-SMAD2/3, αSMA, and cleaved Caspase-3 in D were statistically analyzed after normalization to the band intensity of GAPDH. F The mRNA levels of Smad7, Acta2, Col1a1, and Krt19 in each liver specimen were analyzed by RT–qPCR. The mRNA levels were normalized to those of 18S rRNA. TBG, thyroxine-binding globulin. Scale bar = 20 µm. *p < 0.05, **p < 0.01, ***p < 0.001. All data are the mean ± S.E.M.