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. 2023 Jun 3;14:3236. doi: 10.1038/s41467-023-39050-y

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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. 3 — a Metacore pathway analysis of differentially expressed genes in uninjured PT clusters showing the top 10 significantly affected pathways in γGT-Cre;Tgfbr2^fl/fl compared to Tgfbr2^fl/fl. b Immunoblotting of OXPHOS proteins showing a significant decrease of complex I (NDUFB8) expression in uninjured renal cortices of γGT-Cre;Tgfbr2^fl/fl mice compared to their Tgfbr2^fl/fl littermates; n = 5 (Tgfbr2^fl/fl) and 5 (γGT-Cre;Tgfbr2^fl/fl) mice, p = 0.0041. E-cadherin is used as marker of renal parenchyma and loading control. The dots represent the number of animals per group. c Cell fractionation followed by immunoblotting and quantification of OXPHOS proteins showing a significant decrease of complex I (NDUFB8) basal expression in mitochondria of TβRII^−/− PT cells (n = 3 independent experiments, p = 0.0015). d Bioluminescence measurement of NAD + /NADH ratios showing a decreased relative ratio in TβRII^−/− PT cells (n = 3 independent biological replicates, p = 0.0036). e FACS analysis of DCF-positive cells showing increased basal ROS production in TβRII^−/− PT cells (n = 6 independent experiments, p = 0.0006). f NAD+ treatment significantly decreased ROS in TβRII^−/− PT cells, assessed with DCF and measured by FACS (n = 3 independent biological replicates, p = 0.0027). g NAD+ treatment increased ATP production in TβRII^−/− PT cells to the same level as in TβRII^flox/flox PT cells, measured by a bioluminescence assay (n = 6 independent biological replicates, baseline p = 0.0001 and NAD+ treatment p = 0.4025). h NAD+ treatment decreased lactate production in TβRII^−/− PT cells to the level of TβRII^flox/flox PT cells, measured by a bioluminescence assay (n = 3 independent biological replicates, baseline p = 0.0057 and NAD+ treatment p = 0.3175). Data are presented as mean values ± SEM. Statistical significance was determined by unpaired Student’s t test (two groups) or two-way ANOVA followed by Sidak’s multiple comparisons test with p < 0.05 considered statistically significant. *p < 0.05; **p < 0.01; ****p < 0.0001. Source data are provided as a Source data file.