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Fig. 2. ALS patient-derived fibroblasts display reduced levels of acetylated α-tubulin and disrupted microtubule network. (A) Confocal images of control-derived fibroblasts and ALS patient-derived fibroblasts (E1357K) labeled with antibodies against acetylated α-tubulin (Ac-tub), tyrosinated α-tubulin (Tyr-tub), or α-tubulin (Tub) and DAPI. Scale bar, 5 µm. (B) Western blot analysis of lysates from control and patient fibroblasts using anti-acetylated α-tubulin, anti-tyrosinated α-tubulin, anti-α-tubulin, and anti-GAPDH antibodies. (C and D) Quantitative analysis by densitometric measurements (n=3). The intensities of Ac-tub and Tyr-tub were normalized to that of Tub. Data are presented as mean±SEM. *p<0.001. (E) Confocal images of HeLa cells transiently expressing wild-type GFP-RAPGEF2 (WT) or GFP-RAPGEF2-E1357K (E1357K) labeled with anti-GFP and anti-Actub antibodies. Note that the microtubule network is disrupted in GFP-RAPGEF2-E1357K-expressing cells but not in GFP-RAPGEF2-expressing cells. Scale bar, 5 µm.
Exp Neurobiol 2018;27:550~563
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