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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. 3 — A–C Eight-week-old male Prom1^f/f (f/f) and Prom1^f/f; Krt19-Cre (f/f; Krt19-Cre) mice were subjected to sham surgery (n = 3) or BDL (n = 7–9) for one week. A Each liver specimen was subjected to H&E, Masson’s trichrome and Sirius Red staining, and immunofluorescence analysis of αSMA and CK19. B Liver fibrosis was quantified by measuring the areas stained with Sirius Red. Two to three images of each liver showing Sirius Red staining were obtained. C The mRNA levels of Prom1, Acta2, Col1a1, Krt19, Tgfbr1, and Tgfb1 were determined by RT–qPCR and normalized to those of 18S rRNA. D–F Krt19^CreER; Prom1^+/+; R26^TdTom (Con) and Krt19^CreER; Prom1^f/f; R26^TdTom (f/f) mice were generated for the lineage tracing of CK19-expressing cells. Six-week-old male mice were injected with tamoxifen. Two weeks after injection, the mice were subjected to sham surgery (n = 3) and BDL (n = 7) for 1 week (D). Each liver specimen was analyzed by TdTom fluorescence analysis and CK19 immunofluorescence analysis (E). Statistical analysis of TdTom-expressing cells in E. The TdTom fluorescence area was normalized to DAPI-stained dots (F). Scale bar = 20 µm. All data are the mean ± S.E.M. Con, control.

Fig. 3 — A–C Eight-week-old male Prom1^f/f (f/f) and Prom1^f/f; Krt19-Cre (f/f; Krt19-Cre) mice were subjected to sham surgery (n = 3) or BDL (n = 7–9) for one week. A Each liver specimen was subjected to H&E, Masson’s trichrome and Sirius Red staining, and immunofluorescence analysis of αSMA and CK19. B Liver fibrosis was quantified by measuring the areas stained with Sirius Red. Two to three images of each liver showing Sirius Red staining were obtained. C The mRNA levels of Prom1, Acta2, Col1a1, Krt19, Tgfbr1, and Tgfb1 were determined by RT–qPCR and normalized to those of 18S rRNA. D–F Krt19^CreER; Prom1^+/+; R26^TdTom (Con) and Krt19^CreER; Prom1^f/f; R26^TdTom (f/f) mice were generated for the lineage tracing of CK19-expressing cells. Six-week-old male mice were injected with tamoxifen. Two weeks after injection, the mice were subjected to sham surgery (n = 3) and BDL (n = 7) for 1 week (D). Each liver specimen was analyzed by TdTom fluorescence analysis and CK19 immunofluorescence analysis (E). Statistical analysis of TdTom-expressing cells in E. The TdTom fluorescence area was normalized to DAPI-stained dots (F). Scale bar = 20 µm. All data are the mean ± S.E.M. Con, control.