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. 2024 Aug 14;46(8):8835–8851. doi: 10.3390/cimb46080522

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© 2024 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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The effect of SIRT1 isoforms on mitochondrial function after transfection. HEk293T SIRT1-KO cells were transfected with SIRT1 isoform plasmids and the mitochondrial respiratory profile of these isoforms was examined with the Seahorse XFe96 analyzer. (A) Oxygen consumption rate (OCR) measurements were obtained over time (min) using an extracellular flux analyzer, Seahorse XFe96. (B) Basal mitochondrial OCR of SIRT1 isoform-transfected KO cells was derived by subtracting non-mitochondrial OCR. Maximum OCR was stimulated by FCCP addition. The spare respiratory capacity was calculated as the difference between maximal and basal OCR. (C) Glycolytic rate (ECAR) of SIRT1 isoform-transfected KO cells. (D) A quantitative analysis of the basal and compensatory glycolysis. (E) Quantification of total ATP production rate of KO cells transfected with different SIRT1 isoforms. Graphs represent data normalized with equal cell count, n = 3, one-way ANOVA, Tukey’s post hoc test (ns, p > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

The effect of SIRT1 isoforms on mitochondrial function after transfection. HEk293T SIRT1-KO cells were transfected with SIRT1 isoform plasmids and the mitochondrial respiratory profile of these isoforms was examined with the Seahorse XFe96 analyzer. (A) Oxygen consumption rate (OCR) measurements were obtained over time (min) using an extracellular flux analyzer, Seahorse XFe96. (B) Basal mitochondrial OCR of SIRT1 isoform-transfected KO cells was derived by subtracting non-mitochondrial OCR. Maximum OCR was stimulated by FCCP addition. The spare respiratory capacity was calculated as the difference between maximal and basal OCR. (C) Glycolytic rate (ECAR) of SIRT1 isoform-transfected KO cells. (D) A quantitative analysis of the basal and compensatory glycolysis. (E) Quantification of total ATP production rate of KO cells transfected with different SIRT1 isoforms. Graphs represent data normalized with equal cell count, n = 3, one-way ANOVA, Tukey’s post hoc test (ns, p > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).