Fig. 4
Cholesterol level, membrane fluidity, and lipid rafts-dependent endocytosis are altered in parkin KO MEF cells. a The total cholesterol level was measured in WT and parkin KO MEF cells, as described in ‘Methods’. P values were determined using a Student’s t-test. *p < 0.05. b WT and parkin KO MEF cells were stained with 0.5 μM C-laurdan, and then observed by two-photon microscopy. Images were processed to obtain GP values as described in ‘Methods’. Scale bar indicates 10 μm. c WT and KO MEF cells transfected with siRNA for non-targeting (siCon) and cav-1 (siCav-1) for 48 h, were 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. Scale bar indicates 20 μm. The intensity of three independent experiments was quantified. The mean value of intensity of each experiment was acquired by measuring the intensity of at least five random fields. P values were determined using a Two way ANOVA. **p < 0.01. d Parkin KO MEF cells were transfected with siRNA for non-targeting (SiCon) and cav-1 (SiCav-1) for 48 h, and then the lysates were analyzed by SDS-PAGE and Western blotting