Supplemental Figures

This article contains the following supporting material:

• Supplemental Figure 1.pdf - A series of confocal images showing the morphology of the Golgi apparatus in cells co-transfected with GalT2-YFP plus wt-LIMK1 or LIMK1-kd. Note the condensed appearance of the Golgi apparatus in the neurons co-transfected with wt-LIMK1 and the more extended morphology in those transfected with LIMK1-kd. Note the presence of some small Golgi fragments within the neurites (presumably dendrites) of the neurons transfected with LIMk1-kd. Cultures were transfected 4 days after plating and fixed 18 h later.
  
• Supplemental Figure 2.pdf - (A-C) A sequence of fluorescence images showing the morphology of the Golgi apparatus from a neuron co-expressing galactosyl-transferase T2-EYFP and LIMK1-kd, and visualized 24 h after transfection. Note the fragmentation of the Golgi apparatus. (D-F) Confocal images showing the morphology of the Golgi apparatus after a 3 h treatment with cytochalasin D in a neuron transfected with galactosyl-transferase T2-EYFP (green). The culture was double stained for tubulin (red). Note the fragmentation of the Golgi apparatus. (G-I) Confocal images showing fragmentation of the Golgi apparatus in a neuron co-transfected with LIMK1-kd (red) and galactosyl-transferase T2-EYFP (green) and treated with cytochalasin D for 1 h. Note that LIMK1-kd remains associated with the Golgi fragments. (J-L) Confocal images showing the morphology of the Golgi apparatus in a neuron co-transfected with wt-LIMK1 (red) and galactosyl-transferase T2-EYFP (green) and treated with cytochalasin D for 3 h. Note that the Golgi apparatus remains intact. Bar: 10 μm.
  
• Supplemental Figure 3.pdf - Effect of the expression of HA-tagged Δ-LIM on neuronal morphology. The cells were stained with a rabbit polyclonal antibody against HA (green) and either tyrosinated α-tubulin (red in A) or rhodamine-phalloidin (red in B). The image in A shows the general morphology of a cell overexpressing Δ-LIMK1 12 h after transfection; note the presence of numerous and large growth cone-like structures along the axon The image in B shows a large growth cone from another transfected neuron displaying a very strong fluorescence signal for rhodamine phalloidin. Bars: A, 10 μm; B, 5 μm.
  
• Supplemental Figure 4.pdf - Effect of the expression of HA-tagged LIMK1-kd on neuronal morphology. The cells were fixed 12 h after transfection, stained with a rabbit polyclonal antibody against HA (green) and either a mAb against tyrosinated α-tubulin (red in A, B) or rhodamine-phalloidin (red in E). Cell overexpressing LIMK1-kd display long axonal processes with numerous collateral branches. The growth cones of transfected cells are small and contain less F-actin (small arrows in D) than equivalent ones of non-transfected neurons (long arrows in D). (E) A red-green overlay of the images shown in C and D. Bars: A, B, 5 μm; C-E, 10 μm.
  

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