Mitochondrial Biogenesis Pathway Regulation

The protein expression levels of peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1α), peroxisome proliferator-activated receptor alpha (PPARα), nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (mtTFA) were measured by Western blot (Chen et al., 2018 (link)). The specific methods and steps are shown in Part 2 of the Supplementary Materials. The following antibodies were used: anti-PGC-1α (batch number: ab54481), anti-PPARα (batch number: ab8934), anti-NRF1 (batch number: ab175932), and anti-mtTFA (mitochondrial marker, batch number: ab131607) were provided by Abcam (Cambridge, UK); anti-β-actin (batch number: TA-09) was supplied by Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. (Beijing, China). The mRNA expression levels of PGC-1α, PPARα, NRF1, and mtTFA were measured by real-time PCR (RT-PCR) (Mocker et al., 2019 (link)). The specific methods and steps are shown in Part 3 of the Supplementary Materials. Mitochondrial DNA (mtDNA) was detected using the PCR-fluorescent probe method (Liu et al., 2016 (link)). The specific methods and steps are shown in Part 4 of the Supplementary Materials.