Pe conjugated anti mouse cd40 mhc 2 ccr7 foxp3 pd l1

Recombinant murine granulocyte–macrophage colony stimulating factor (rmGM-CSF), recombinant murine interleukin-4 (rhIL-4) and murine MIP-3β (CCL19) were purchased from Peprotech (Rocky Hill, NJ, USA). LPS and FITC-conjugated dextran (molecular weight: 43 kDa) were purchased from Sigma‒Aldrich (St. Louis, MO, USA). Metformin was purchased from Medchem Express (New Jersey, USA). FITC-conjugated anti-mouse CD80/CD86/CD4, PE-conjugated anti-mouse CD40/MHC-II/CCR7/Foxp3/PD-L1, and APC-conjugated anti-mouse CD11c/CD25 antibodies were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Mouse IL-12p40, IL-6, TNF-α, IFN-γ, IL-10 and TGF-β enzyme-linked immunosorbent assay (ELISA) kits were obtained from 4A Biotech Co., Ltd (Beijing, China). Antibodies against phosphorylated forkhead box-O3a (pFoxO3a) (s253) and Foxo3a were obtained from Huabio Co., Ltd. (Hangzhou, China). Antibodies against GAPDH, TRITC phalloidin, 40,6-diamidino-2-phenylindole (DAPI) and Cell Counting Kit 8 (CCK-8) were obtained from Solarbio (Beijing, China). A mouse naïve CD4⁺ T-cell isolation kit was purchased from Miltenyi Biotec (Cologne, Germany). Hematoxylin and eosin (H&E) staining solution was obtained from Thermo Fisher Scientific (Waltham, MA, USA).

Recombinant murine granulocyte–macrophage colony stimulating factor (rmGM-CSF), recombinant murine interleukin-4 (rhIL-4) and murine MIP-3β (CCL19) were purchased from Peprotech (Rocky Hill, NJ, USA). LPS and FITC-conjugated dextran (molecular weight: 43 kDa) were purchased from Sigma‒Aldrich (St. Louis, MO, USA). Metformin was purchased from Medchem Express (New Jersey, USA). FITC-conjugated anti-mouse CD80/CD86/CD4, PE-conjugated anti-mouse CD40/MHC-II/CCR7/Foxp3/PD-L1, and APC-conjugated anti-mouse CD11c/CD25 antibodies were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Mouse IL-12p40, IL-6, TNF-α, IFN-γ, IL-10 and TGF-β enzyme-linked immunosorbent assay (ELISA) kits were obtained from 4A Biotech Co., Ltd (Beijing, China). Antibodies against phosphorylated forkhead box-O3a (pFoxO3a) (s253) and Foxo3a were obtained from Huabio Co., Ltd. (Hangzhou, China). Antibodies against GAPDH, TRITC phalloidin, 40,6-diamidino-2-phenylindole (DAPI) and Cell Counting Kit 8 (CCK-8) were obtained from Solarbio (Beijing, China). A mouse naïve CD4⁺ T-cell isolation kit was purchased from Miltenyi Biotec (Cologne, Germany). Hematoxylin and eosin (H&E) staining solution was obtained from Thermo Fisher Scientific (Waltham, MA, USA).