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Fig. 8. Co-localization and immunoprecipitation of Cx43 and P2X₇ proteins in peritoneal macrophages. (A) Cx43 immunolabeling, (B) P2X₇ immunolabeling and (C) TOPRO3 staining showing the nuclei of the J774 cells. Bar, 20 µm. (D) Reconstruction of 49 optical slices of 0.8 µm thickness. The green line indicates an orthogonal section from (D), projected laterally in (E); the red line indicates an orthogonal section from (D), projected laterally in (F); the blue line indicates the level of the 25th optical slice in (D). Confocal microscopy demonstrates co-localization of the Cx43 and P2X₇ proteins (yellow) to the membrane of the macrophages (D-F, arrows). Notice the Cx43 (green) and P2X₇ (red) specific labeling located at membrane apposition areas (A,B, arrows). (G) Lane 1 shows a 69 kDa band corresponding to P2X₇ receptor obtained after immunoprecipitation of peritoneal macrophage lysates with Cx43 antibodies, and lane 2 shows the corresponding band obtained from whole peritoneal macrophage cell lysates probed with the P2X7 antibody.

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