Slc25a1

RAW264.7 cells and kidney tissues were collected and added with RIPA lysis buffer (Beyotime, Shanghai, China). After thorough lysis on ice, the lysates were centrifuged at 4°C, and the supernatants were collected. The concentration of total proteins in the supernatant was determined using the BCA protein assay kit (Solarbio, Beijing, China). Equal amounts of protein were separated using SDS-PAGE and transferred onto PVDF membranes (Millipore Sigma, Burlington, MA, USA). The membranes were blocked at room temperature for 60 minutes and then incubated overnight at 4°C with the corresponding primary antibodies. Afterwards, the corresponding species-specific secondary antibodies (Proteintech, Wuhan, China) were incubated on a shaker at room temperature for 60 minutes. Finally, ECL Western Blot Substrates (Zenbio, Chengdu, China) were applied to the protein bands, and chemiluminescent signals were detected using imaging equipment. All antibodies used for Western blot analysis are as follows: β-actin (20,536-1-AP, Proteintech), TNF-α (17,590-1-AP, Proteintech), IL-6 (12,912, Cell Signaling Technology), SLC25A1 (15,235-1-AP, Proteintech), HK2 (22,029-1-AP, Proteintech), PFKFB3 (13,763-1-AP, Proteintech), PKM2 (15,822-1-AP, Proteintech), NDUFB8 (14,794-1-AP, Proteintech), UQCRFS1 (18,443-1-AP, Proteintech), ACLY (ab40793, Abcam), HIF-1α (ab179483, Abcam).