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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Cell Immunol. 2012 Oct 1;279(1):46–52. doi: 10.1016/j.cellimm.2012.09.004

Figure 2. NKLAM is localized to the macrophage phagosome.

Figure 2

(A) Adherent wild type BMDM were incubated with fluorescent red E. coli (left panel) for 60 min at 37°C. The monolayers were fixed in paraformaldehyde and labeled with anti-NKLAM (green, middle panel). Confocal photomicrographs taken at the midpoint of the z series were overlaid (right panel) and processed with the ImageJ Colocalization plugin. Arrows (inset) depict areas of specific colocalization, which are denoted in white. (B) RAW264.7 were untreated (−) or stimulated (+) with LPS/IFNγ for 18hr at 37°C. The cells were then incubated with IgG-opsonized magnetic latex beads for 45 min. The phagosomes were isolated as described in the Materials and Methods section. Equal amounts of phagosome protein were immunoblotted for NKLAM and phagosome markers LAMP1 and EEA1. (C) LPS/IFNγ-stimulated WT and NKLAM-deficient (KO) BMDM were used in experiments that were identical to (B).