740 y p hy p0175

Melatonin (HY-B0075) and 740 Y-P (HY-P0175) were bought from MedChemExpress (MCE, China). A 1mol/L stock solution was attained by dissolving melatonin in dimethyl sulfoxide (DMSO), which was then kept at −20°C in the dark. The DMSO and N-acetyl-L-cysteine (NAC) were provided by the Beyotime (Beyotime Institute of Biotechnology, Shanghai, China). The Cell Counting Kit-8 (CCK-8) was bought from US EVERBRIGHT INC. The primary antibodies used in this study included Bax (ab32503, Abcam, Cambridge, MA, USA), Bcl-2 (ab32124, Abcam, Cambridge, MA, USA), Cytochrome C (ab76237, Abcam, Cambridge, MA, USA), Cleaved Caspase-3 (9662, Cell Signaling Technology Inc, CST, MA, USA), PI3K (AB3263, Technology, Shanghai, China), phospho-PI3K (CY6427), phospho-Akt (AY0421), and Phospho-mTOR (CY5996). The HRP-linked goat anti-mouse and anti-rabbit secondary antibodies were also bought from CST.

Anti-mouse CD3e (100340), anti-mouse CD28 (102116), recombinant mouse IL-2 (575406), recombinant mouse IL-6 (575702), recombinant mouse TGF-β (763102), recombinant mouse IL-12 (577004), recombinant mouse IL-23 (589002), Brilliant Violet 421-conjugated anti-mouse CD4 (100443), PerCP anti-mouse CD8a (100731), PE-conjugated anti-mouse/human CD44 (103008), APC-conjugated anti-mouse CD62L (104412), PE/Cyanine7-conjugated anti-mouse IFN-γ (505826), Brilliant Violet 421-conjugated anti-mouse IL-17A (512321), PE-conjugated anti-mouse IL-17A (506904), Alexa Fluor^® 647 anti-mouse/rat/human FOXP3 (320014), FITC anti-mouse CD11c (117306), Brilliant Violet 421™ anti-mouse I-A/I-E (107620), PE/Cy7 anti-mouse CD86 (105014), and PE anti-mouse F4/80 (123110) were purchased from Biolegend (San Diego, CA, USA). Anti-phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) (4228S), anti-PI3 Kinase p85 (4292S), anti-phospho-AKT (Ser473) (D9E) XP^® Rabbit mAb (4060S), anti-AKT (9272S), anti-PKM2 (4053S) and anti-β-Actin (3700S) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA); anti-PGK1 (A12686), anti-ENO1 (A11448), and anti-LDHA (A1146) antibodies were got from Abclonal (Wuhan, China); anti-HK2 (22029-1-AP) and anti-Glut1 (21829-1-AP) antibodies were obtained from Proteintech (Wuhan, China); and 740 Y-P (HY-P0175) was ordered from Medchem Express (Shanghai, China).

AMG 510, the MEK inhibitor GSK1120212 (trametinib), the PI3K inhibitor GDC-0941 (pictilisib), and the PI3K agonist 740 Y-P (HY-P0175) were purchased from MedChemExpress, the STAT3 inhibitor Stattic was purchased from Selleck, and the MAPK agonist PAF C-16 (sc-201009) was purchased from Santa Cruz Biotechnology. Unless otherwise noted, all chemicals were purchased from MilliporeSigma.