Fig. 5 | Nature Communications

Fig. 5

From: Membrane-cytoskeletal crosstalk mediated by myosin-I regulates adhesion turnover during phagocytosis

Fig. 5

Actin waves in dKO BMDM have higher density of branched actin filaments. a Immunostaining of Arp2/3 complex in WT and dKO BMDM during frustrated phagocytosis. Cells were counter-stained with fluorescently labeled phalloidin. Zoom of boxed regions shown on the right. Scale bar, 10 μm; zoom panel scale bar, 1 μm. b Treating dKO cells with low doses of Arp2/3 inhibitor CK-666 partially rescues actin wave morphology. Graph depicting the mean percentage of cells forming diffuse or aggregated actin waves of WT and dKO BMDM in the presence of 1 μM CK-666. Data from two independent experiments (n ≥ 40 cells per genotype, judged blindly). cg Correlative confocal and platinum replica EM of actin wave in a representative WT macrophage. c Confocal section of a WT cell stained with fluorescently labeled phalloidin for correlative fluorescence image. Scale bar, 5 µm. d Platinum replica EM image of the macrophage shown in c. Scale bar, 5 µm. e Overlay of the enlarged confocal (green) and platinum replica EM (gray) images corresponding to the boxed region in d. Scale bar, 0.5 µm. f, g Sequential magnifications of boxed regions in e and f, respectively, showing supramolecular architecture of individual phagocytic adhesions. Scale bars, 0.2 µm. hl Correlative confocal and platinum replica EM of actin wave in a representative dKO macrophage. h Confocal section of a dKO cell stained with fluorescently labeled phalloidin for correlative fluorescence image. Scale bars, 5 µm. i Platinum replica EM image of the macrophage shown in h. Scale bars, 5 µm. j Overlay of the enlarged confocal (green) and platinum replica EM (gray) images corresponding to the boxed region in i. Scale bars, 0.5 µm. k, l Sequential magnifications of boxed regions in j and k, respectively, showing unbridled actin polymerization. In l, image contrast is changed by adjusting gamma. Scale bars, 0.2 µm. See also Supplementary Fig. 9 and Supplementary Movie 12

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