FIGURE

Fig. 5

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ZDB-FIG-191224-4
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Villani et al., 2019 - Clearance by Microglia Depends on Packaging of Phagosomes into a Unique Cellular Compartment
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Fig. 5

Molecular and Ultra-Structural Characterization of the Gastrosome

(A–C) Fixed RAW264.7 macrophages co-cultured with HK H2B-mCherry (red) and HeLa H2B-GFP (green) for 3 h (A), 6 h (B), and 9 h (C) after induction of apoptosis in HeLa and HK. The yellow arrow indicates the collection compartment (gastrosome).

(D) Schematic representation of the experiment in (A)–(C).

(E) RAW264.7 macrophages fixed 9 h after apoptosis induction of unlabeled HeLa cells. Immunostaining against Lamp1 (red). The white arrow indicates the collection compartment (gastrosome). Scale bar, 5 μm.

(F) RAW264.7 macrophages fixed 9 h after apoptosis induction of unlabeled HeLa cells. Immunostaining against Cathepsin D (red). The white arrow indicates the collection compartment (gastrosome). Scale bar, 5 μm.

(G–I) CLEM of RAW264.7 macrophages fixed 9 h after induction of apoptosis in HK H2B-mCherry (red) and HeLa H2B-GFP (green). (G) Confocal microscopy data. Scale bar, 5 μm.

(H) Low magnification EM data. White arrowhead points to the compartment of interest. Scale bar, 5 μm. (I) High magnification of the compartment of interest in TEM data. Scale bar, 2 μm.

(J) Segmentation of μCT data. 4 dpf zebrafish embryo outline is in blue. Yellow and red mark near-infra-red-branding as detected by 2-photon microscopy and μCT, respectively. Outline of the microglial cell of interest is in green.

(K) Segmentation of the cell of interest. Green outlines the cell body. Blue marks the nucleus. Phagosomes are in red. The receiving compartment (gastrosome) is in yellow. See also Video S7.

(L) Higher magnification of the cell of interest. Green outlines the cell body, blue marks the nucleus, phagosomes are in red, and yellow marks the compartment of interest (gastrosome). White arrowhead points to the compartment of interest. Scale bar, 5 μm.

(M) High magnification of the gastrosome in SBF-SEM data. P indicates a phagosome. Scale bar, 2 μm.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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Reprinted from Developmental Cell, 49(1), Villani, A., Benjaminsen, J., Moritz, C., Henke, K., Hartmann, J., Norlin, N., Richter, K., Schieber, N.L., Franke, T., Schwab, Y., Peri, F., Clearance by Microglia Depends on Packaging of Phagosomes into a Unique Cellular Compartment, 77-88.e7, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell