Phrodo green intracellular ph indicator

hiPSC-RPE cells were seeded at different conditions and cultured overnight. pHrodo Green Intracellular pH Indicator (Thermo Fisher Scientific, Waltham, MA, USA) was added to the RPE cells to observe the phagocytosis-induced intracellular pH change, according to the manufacturer’s instruction. After incubation at the indicated concentrations and time, cells were collected and fixed. FITC signal intensity was measured during flow cytometry analysis.

PE-labeled anti-human SLAMF7 antibody (clone, 162.1) and PE-labeled isotype control were purchased from Biolegend. APC-labeled anti-CD3, FITC-labeled anti-CD19 and APC-labeled anti-CD47 were purchased from Immunotools (Germany). FITC-labeled anti-CD20 was purchased from Thermofisher. Alexa594-labeled CD11b was purchased from Biolegend. Anti-human CD47 IgG4 antibody (Inhibrix, clone B6H12) was generated in-house by Aduro Biotech Europe (ABE). F(ab′)2 of Inhibrix was prepared with Pierce F(ab′)2 preparation kit. F(ab′)2 generation was confirmed by staining for human IgG4 (Supplementary Figure 1D). Cell proliferation dye V450 was purchased from Thermofisher. Phrodo Green pH indicators was purchased from Thermofisher. Lymphoprep was purchased from Axis-Shield PoC AS, Norway. pHrodo Green intracellular pH indicator was purchased from ThermoFisher. RNeasy mini kit was purchased from Qiagen. iScript cDNA Synthesis Kit was purchased from Bio-Rad.