Cpi 613

To investigate the origin of redox signals, various inhibitors such as sodium diethyldithiocarbamate, GKT136901 (Sigma‐Aldrich, MO, USA), Mito‐tempo (MedChemExpress, NJ, USA), FCCP, oligomycin, rotenone and antimycin A (Agilent, CA, USA), and Batimastat (Selleckchem, Seoul, South Korea) were used as inhibitors. All of the inhibitors were incubated for 24 h at 37 °C. Furthermore, the cells were treated with CPI‐613 (MedChemExpress, NJ, USA) for drug screening and incubated for 24 h at 37 °C. After treatment, the cells were washed with DPBS for EC detection within 30 s and cytotoxicity testing.

Dulbecco’s modified eagle’s medium (DMEM), glutamine-free DMEM, glutamine, IL-4 and LPS were purchased from Sigma Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) was obtained from PAN Biotech (Bavaria, Germany). CPI-613, MC3482 and succinyl phosphonate (SP) were obtained from MedChemExpress (Monmouth Junction, NJ, USA). BLZ945 and L-α-aminoadipic acid (L-α-A) were purchased from APExBIO Technology LLC (Houston, TX, USA). CB-839 was obtained from Selleck Chemicals (Houston, TX). Co-immunoprecipitation (Co-IP) assay kit was purchased from Thermo Fisher Scientific (Rockford, IL, USA). Primary antibodies against arginase-1 (Arg1), CD206, PDH E1 component subunit alpha (PDHA1), phosphorylated PDHA1 (p-PDHA1), tubulin and IgG isotype control antibodies were purchased from Abcam (Cambridge, UK). Primary antibodies against pan-succinyllysine (Pan-succK) and pan-acetyllysine (Pan-acetK) were obtained from PTM BIO (Hangzhou, China). Primary antibodies against sirtuin SIRT5 and F4/80 and HRP-linked anti-mouse and anti-rabbit secondary antibodies were purchased from Cell Signaling Technology (Boston, MA, USA). Primary antibody against proliferating cell nuclear antigen (PCNA) was purchased from Proteintech Group (Rosemont, IL, USA). FITC and Cy3- labelled secondary antibodies were purchased from Beyotime Biotechnology (Shanghai, China).