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. 2004 Jul;15(7):3433–3449. doi: 10.1091/mbc.E03-05-0328

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Copyright © 2004, The American Society for Cell Biology

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Figure 8. — LIMK1 promotes the accumulation of Par3/Par6 at axonal growth cones and waves. (A and B) Confocal micrographs showing the distribution of HA-tagged LIMK1 (red) and Par3 (green) in cultured hippocampal pyramidal neurons. Cells were transfected with wt-LIMK1 2 h after plating, fixed 18 h later and stained with a mouse antibody against HA and a rabbit polyclonal antibody against Par3 (diluted 1/200). (C and D) Confocal micrographs showing the distribution of HA-tagged LIMK1 (green), F-actin (red), and Par6 (blue) in cultured hippocampal pyramidal neurons. Cells were transfected as described previously. Note that only the transfected neurons display intense Par3/Par6 immunofluorescence at the axonal growth cone (arrows). (F and G) Confocal micrographs showing colocalization and accumulation of HA-tagged LIMK1 (red) and Par3 (green) within an axonal shaft and a wave (arrows). (H and I) Confocal micrographs showing colocalization and accumulation of HA-tagged LIMK1 and transfected Par3 within the terminal axonal arbor of a cultured hippocampal pyramidal neuron. (J) Red-green overlay of the images shown previously. (K) Confocal image showing the axonal shaft of a neuron transfected with Par3 alone. For these experiments cultures were transfected 1 d after plating and fixed 18 h later; the Par3 antibody was used at a dilution of 1/500.