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. 2023 May 25;14:3007. doi: 10.1038/s41467-023-38771-4

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© The Author(s) 2023

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. 4 — a Cell cycle analysis by flow cytometry for HK-2 in different groups. (n = 6 biologically independent experiments). b Representative Western blot gel documents and summarized data showing the ratio of cyclin B1 to cyclin D1 densities standardized to β-actin and protein levels of p21 in HK-2 with AA treatment. (n = 6 biologically independent experiments). c The level of TGF-β1 in the culture supernatant from HK-2 treated with AA for 24 h. (n = 6 biologically independent experiments). d Experimental scheme for cell treatment: after 48 h treatment of HK-2 cells with aristolochic acid, the drug was washed out and the cells continued in culture for 24 h. The conditioned medium was then collected and added to serum-starved NRK-49F cells. e Protein levels of Vimentin and PCNA in fibroblasts treated with conditioned medium from control or G2/M-arrested HK-2 cells. (n = 6 biologically independent experiments). f Representative photomicrographs of coimmunostaining with antibodies to Ki-67 (anti–Ki-67) and p-H3 (anti–p-H3) on kidneys and the percentage of Ki-67⁺ p-H3⁺ cells among total Ki-67⁺ tubular cells in different groups. Scale bar: white = 20 μm. (n = 6 mice per group). g Representative Western blot gel documents and summarized data showing the ratio of cyclin B1 to cyclin D1 densities standardized to β-actin in isolated tubules and protein levels of TGF-β1 in cortex of kidney from different groups. (n = 6 mice per group). h Photomicrographs and quantifications showing PCNA expression in kidney from different groups of mice (up panel). PCNA-positive cells per high power field (hpf) are counted and shown. Scale bar: black = 50 μm. Representative photomicrographs of kidney sections stained for α-SMA, PDGFRβ⁺, and DAPI (down panel), as well as quantitative analysis of a-SMA staining in kidney. Scale bar: white = 20 μm. (n = 6 mice per group). Data are expressed as mean ± SEM (a–c, e–h). Two-way ANOVA followed by Tukey’s post-test (a–c, e–h). Source data are provided as a Source Data file.