Monensin sodium salt

The human neuroblastoma SH-SY5Y (ATCC^®-: CRL-2266™) cell line was used in this study. Cells were incubated in DMEMF12 (Dulbecco’s Modified Eagle Medium) culture containing penicillin (20 units/mL), streptomycin (20 μg/mL), and 10% fetal bovine serum at 37 °C under 95% air and 5% CO₂ pressure. Monensin sodium salt, Na⁺ ionohore (Figure 1) (Abcam, cat. no.: ab120499, Boston, MA, USA), and rap (Figure 1) (MedChemEkspress, cat. no.: HY-10219, Newark, NJ, USA) were dissolved in ethanol.

Monensin sodium salt, Na⁺ ionophore (Abcam, cat. no: ab120499, Boston, MA, USA), and rapamycin (MedChemEkspress, cat. no: HY-10219, Newark, NJ, USA) were dissolved in ethanol.

For cell-surface analysis, cells were stained with anti-mouse Zombie Violet™ Fixable Viability Kit (423114), Zombie NIR™ Fixable Viability Kit (423106), CD45 (103114), CD11b (101205), CD86 (105012), CD4 (100408), CD8a (100752), Gr-1 (108411), and PD-L1 (124311, 124308) in recommended antibody concentrations and incubated at 4 °C for 30 min. For the T-cell intracellular IFN-γ (505808) cytokine staining, cells were fixed and permeabilized after stimulation with Phorbol 12-myristate 13-acetate (PMA) (ab120297, Abcam, 100 ng/mL), Monensin sodium salt (ab120499, Abcam, 1 ug/mL), and Ionomycin calcium salt (5608212, PeproTech, 100 ng/mL) for 6 h. For the CD206 (141706) and Foxp3 (126408) staining, cells were also fixed and permeabilized. All flow cytometry antibodies were purchased from Biolegend (San Diego, CA, USA).