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Fig. 7. Influence of the microtubule cytoskeleton on peroxisomal morphology and dynamics after silencing of DLP1. COS-7 cells stably expressing GFP-PTS1 (GFP) were transfected with (A) buffer (Con) or (B-G) DLP1 siRNA duplexes (siRNA) and processed for immunofluorescence (A-D) or live-cell imaging (E-G). Cells in C,D,G were treated with nocodazole to depolymerize microtubules. (A,B,D) Immunofluorescence of COS-7 cells using antibodies to tubulin (Tub). (C) Visualization of peroxisomes labeled with GFP-PTS1 after transfection with DLP1 siRNA and treatment with nocodazole. Arrows point to peroxisomal aggregates. (E-G) Images of the motile behavior of individual, elongated peroxisomes in vivo after silencing of DLP1. Numbers in the top right of the panels show time elapsed in seconds. In E, an elongated peroxisome (arrow) moves to the right of the image (0 to 30 seconds) before it continues to move on a circular track (50 to 65 seconds). In F, an elongated peroxisome (arrows) with a globular structure at one end (*) moves from the left to the right of the image (0 to 5 seconds). Afterwards it emanates a tubular projection and moves to the left of the image (30 seconds), presumably to interact with another peroxisome, before it retracts and collapses into a circular structure (65 seconds). In G, the dynamic behavior of elongated peroxisomes after depolymerization of microtubules with nocodazole is shown. Arrows mark the positions of elongated peroxisomes and arrowheads point to segmented organelles. N, nucleus. Bars, 10 µm (A-D), 5 µm (E-G).
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