TrkAIII Promotes Microtubule Nucleation and Assembly at the Centrosome in SH-SY5Y Neuroblastoma Cells, Contributing to an Undifferentiated Anaplastic Phenotype
Figure 3
Representative IF images and histograms of MT regrowth assays, demonstrating (a) indirect IF changes in α-tubulin positive MT regrowth (red) from the γ-tubulin positive centrosome (green) in untreated and CEP-701-treated (100 nM) TrkAIII SH-SY5Y transfectants and in untreated TrkAI SH-SY5Y transfectants at 0, 5, and 15 minutes after nocodazole washout. (b) Histogram demonstrating the differences in mean (±S.E.) α-tubulin IF intensity in untreated and CEP-701-treated (100 nM for 1 hour) TrkAIII SH-SY5Y transfectants and in untreated TrkAI and control pcDNA SH-SY5Y transfectants at 0, 5, and 15 minutes after nocodazole washout, normalised with respect to untreated TrkAIII transfectants (arbitrary value 100%; per group; * = statistical significance). (c) Indirect IF demonstrating differences in α-tubulin positive MT regrowth area (red) in untreated and CEP-701-treated (100 nM) TrkAIII SH-SY5Y transfectants and in untreated TrkAI SH-SY5Y transfectants at 0, 5, and 15 minutes after nocodazole washout. (d) Histogram demonstrating differences in α-tubulin positive MT regrowth area in untreated and CEP-701-treated (100 nM) TrkAIII SH-SY5Y transfectants and in untreated TrkAI and control pcDNA SH-SY5Y transfectants at 0, 5, and 15 minutes after nocodazole washout, normalised with respect to untreated TrkAIII transfectants (arbitrary value of 100%; per group; * = statistical significance). (e) Histogram demonstrating the differences in γ-tubulin positive centrosome size in untreated control, TrkAI, and TrkAIII SH-SY5Y transfectants, normalised with respect to untreated control transfectants given the arbitrary value of 1 ( per group; * = statistical significance).
