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. 2007 Apr 2;117(4):919–930. doi: 10.1172/JCI30328

Figure 8. Plekhm1 colocalizes to late endosomes/lysosomes with Rab7 in prefusion osteoclasts.

Figure 8

(A–D) Prefusion human osteoclasts were transfected with Plekhm1-EGFP or EGFP-Rab7 and then analyzed 24 hours later by confocal microscopy. We also labeled acidic vesicles with lysotracker red (A), late endosomes/lysosomes with TRITC-dextran (B), and recycling endosomes with Alexa Fluor 633–transferrin (C) prior to fixation. Plekhm1 was associated mainly with intracellular vesicles that accumulated lysotracker and endocytosed dextran (A and B, arrows) but not transferrin, indicating that plekhm1 is localized to late endosomes/lysosomes. (D) Osteoclast-like cells were counterstained with an antibody to the VNR (red stain) and the nuclei stained with TO-PRO-3 iodide (blue stain). Both EGFP-Rab7 and Plekhm1-EGFP were localized to intracellular vesicles in VNR-positive osteoclasts. (E) Prefusion osteoclast cells were cotransfected with Plekhm1-dsRedM and EGFP-Rab7, then stained with an anti-VNR antibody. Plekhm1 and Rab7 almost completely colocalize on intracellular vesicles in VNR-positive prefusion osteoclast cells. Panels represent 1-μm xy optical sections. Scale bars: 5 μm.