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. Author manuscript; available in PMC: 2017 Oct 18.
Published in final edited form as: Immunity. 2016 Oct 18;45(4):861–876. doi: 10.1016/j.immuni.2016.09.014

Figure 3. Granuloma Epithelialization Occurs within Macrophages and Involves Multiple Adhesive Pathways.

Figure 3

(A) E-cadherin was detected by immunofluorescence (red) and colocalized with endogenous Gt(jup-citrine)ct520a (green) in granulomas in 2 wpi M. marinum (cyan) infected animals. Scale bar – 50 µm. (B) E-cadherin staining (red) and plakoglobin fluorescence (green) within cell populations that are positive for the pan-leukocyte marker l-plastin (magenta) in granulomas from 2 wpi animals. Scale bar – 25 µm. (C) Diagram of lineage tracing approach used to label macrophages. (D) Images of tdTomato expression in animals expressing either macrophage-specific Cre alone (Tg(mfap4:icre-p2a-tomato)) or double transgenic animals in which macrophage specific Cre rearranges the blue-to-red lineage tracing cassette (Tg(ubb:BSR; mfap4:icre)), resulting in robust red fluorescence from the reporter cassette. Asterisks mark autofluorescent melanocytes. Scale bar – 100 µm. (E) A necrotic granuloma in a 2 wpi Tg(mfap4:icre; ubb:BSR); Gt(jup-citrine)ct520a animal infected with 400 FB of cerulean-tagged mycobacteria. Images show macrophages (red) localizing to plakoglobin positive adherens junctions (green); infecting mycobacteria are visualized in cyan. Yellow box indicates magnified areas shown in the row below. Images are representative of granulomas from 12 animals. Scale bar – 50 µm. (F) Electron microscopy images of desmosomes (yellow box), adherens junctions (green box) and tight junctions (red box) in granuloma macrophages in M. marinum infected 2 wpi Tg(mfap4:icre; ubb:BSR) zebrafish. (G) Image of desmosomes in 2 wpi Tg(mfap4:icre; ubb:BSR) zebrafish. Note in inset the filament network associated with desmosomes. See also Figure S2 and S3.