Figure 6
Effects of Syk-KO on the expression of actin-binding proteins. (a) Representative images of immunofluorescence staining of macrophage-like differentiated parental and Syk-KO HL60 cells using antibodies against PLCγ2 (green) after incubation with complement-opsonized Texas Red-labeled zymosan particles for 15 min. Scale bars show 10 μm. (b) Immunoblotting of Syk, PLCγ2, Cofilin, Vinculin, and α-tubulin in parental and Syk-KO HL60 cells (left; non-differentiated, right; macrophage-like differentiated). The original images of the immunoblotting analysis are presented in Supplementary information. (c) Representative images of F-actin (Alexa Fluor 488-labeled phalloidin) during phagocytosis of macrophage-like differentiated parental and Syk-KO HL60 cells after incubation with complement-opsonized Texas Red-zymosan particles for 15 min, followed by incubation for additional 1 h in the presence or absence of U-73122. Arrows show the phagosomes that are not surrounded by F-actin and arrowheads show the phagosomes surrounded by F-actin. Scale bars shows 10 μm. (d) Quantification of the percentage of the phagosomes surrounded by F-actin in macrophage-like differentiated parental and Syk-KO HL60 cells during phagocytic process in the presence or absence of U-73122 (1 h and 2 h). More than 150 phagosomes per single condition were analyzed. Data show the means ± SD derived from three independent experiments. p values were calculated using a two-tailed unpaired Student’s t-test. ** < 0.01.
