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. 2022 Aug 27;13(8):739. doi: 10.1038/s41419-022-05191-z

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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. 1 — A Left panel: fold changes (relative to normoxia) in mitochondrial superoxide levels (% MitoSOX⁺ cells) for PTEC cultured under normoxic and hypoxic conditions. Bar graphs represent mean ± standard error mean (SEM). Symbols represent individual donor PTEC; n = 5. *P < 0.05, paired t test. Right panel: representative MitoSOX staining (black unfilled) compared with unstained control (grey filled) for PTEC cultured under normoxic and hypoxic conditions. Mitochondrial superoxide levels (% MitoSOX⁺ cells) are presented for each histogram, with fold change (FC) value relative to normoxic PTEC also shown. B Left panel: Fold changes (relative to normoxia) in mitochondrial membrane potential [measured as ratio of ΔMFI JC-1 red/ΔMFI JC-1 green; with delta median fluorescence intensity (ΔMFI) representing MFI test – MFI unstained control] for PTEC cultured under normoxic and hypoxic conditions. Bar graphs represent mean ± SEM. Symbols represent individual donor PTEC; n = 4. *P < 0.05, paired t test. Right panel: representative JC-1 dot plots of PTEC cultured under normoxic and hypoxic conditions. Mitochondrial membrane potential (ΔΨ_mt) values are presented for each histogram, with fold change (FC) value relative to normoxic PTEC also shown. C Left panel: fold changes (relative to normoxia) in γ-H2AX protein levels (as a ratio of loading control β-tubulin) for PTEC cultured under normoxic and hypoxic conditions. Bar graphs represent mean ± SEM. Symbols represent individual donor PTEC; n = 4. *P < 0.05, paired t test. Right panel: western blot for γ-H2AX for PTEC cultured under normoxic and hypoxic conditions (15 µg total protein per lane). Representative images from one of four donor PTEC are presented; full and uncropped western blot available as Supplementary Material. D, E Fold changes (relative to normoxia) in cell viability (MTT assay) (D) and HMGB1 (ELISA) (E) for PTEC cultured under normoxic and hypoxic conditions. Bar graphs represent mean ± SEM. Symbols represent individual donor PTEC; n = 7 for viability data and n = 3 for HMGB1 data. **P < 0.01, paired t test.