Supplementary Figure 7: Substrate-specific modifiers of phagocytosis.
From: Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens
Assay of U937 cell lines expressing sgRNAs that are simultaneously challenged with two phagocytic substrates within the same well, one substrate labeled with pHrodo green and the other with pHrodo red. Phagocytosis of both substrates is measured over time using live-cell microscopy. a, Ratio of the total red area (indicating zymosan phagocytosis) to the total green area (indicating 1.3-μm bead phagocytosis) at the time point at 5 h of the phagocytosis assay in cells expressing control sgRNAs (gray) or sgRNAs targeting ITGB2, TLN1, FERMT3 and PLEK (blue). Values represent mean ± s.e.m. of n = 4 replicate wells. Representative of three independent experiments. b, Ratio of the total red area (indicating C3b-opsonized RBC phagocytosis) to the total green area (1.3-μm bead phagocytosis) at the time point at 5 h of the phagocytosis assay in cells expressing control sgRNAs (gray) or sgRNAs targeting ITGB2, TLN1, FERMT3, and PLEK (blue). Values represent mean ± s.e.m. of n = 4 replicate wells. Representative of two independent experiments. c, Phagocytosis of pHrodo-labeled amyloid-β (Aβ) aggregates over time in differentiated U937 cells expressing control sgRNAs (black and gray), a clonal NCKAP1L KO U937 line (light blue), a clonal NHLRC2 KO U937s line (dark blue), and a U937 line expressing a control sgRNA with the actin polymerization inhibitor cytochalasin D (red). Values represent mean ± s.e.m. of n = 4 replicate wells. Representative of three independent experiments.
