Fig. 2. UCH-L1 regulates lipid raft-dependent endocytosis. (A) Western blot was performed using lysates of NT and UCH-L1 KD #1 and #2 SH-SY5Y cells. (B) The cells were lysed in ice-cold 1% Triton X-100 buffer and fractionated as soluble or insoluble fractions, and then the samples were analyzed by Western blot. (C) The cells were incubated with 50 nM BOIPY® FL C5-Lactosylceramide and 2.5 µg/ml rhodamine-conjugated transferrin for the indicated times. (D) SH-SY5Y cells were treated with LDN-57444 for 24 h, and then incubated with 50 nM BOIPY® FL C5-Lactosylceramide and 2.5 µg/ml rhodamine-conjugated transferrin for the indicated times. The cells were then fixed and observed by confocal microscopy. The intensity was analyzed with the ImageJ program. Values are representative of three independent experiments. Scale bar indicates 20 µm. *p<0.05, **p<0.01 against control by one-way ANOVA.
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