Fig. 1.

Reduced GCase activity does not influence α-syn aggregation in neonatal mice. (A) The wt mice at P8 were i.p. injected with CBE at 100 mg/kg for 7 d, and cortex was analyzed for GCase activity and GSLs substrates (GluCer and GluSph). Galactosylceramide (GalCer) was analyzed as a specificity control. Lipids were normalized to inorganic phosphate (Pi). (B) Whole brains were sequentially extracted and separated into triton X-100 soluble (T-sol) and insoluble (T-insol) fractions followed by Western blot for α-syn. β-iii-tubulin and Coomassie Brilliant Blue (CBB) were used as a loading controls. Quantifications are shown on the right normalized to CBB. C20 (rabbit) and syn505 (mouse) were sequentially probed on the same blot using two different, fluorescent secondary antibodies. (C) T-sol lysates were analyzed by SEC/Western blot to reveal the levels of HMW oligomers and low–molecular weight (LMW) monomers of α-syn. CBB, β-iii-tubulin, and GAPDH are loading controls. Quantification is on the right. Values are the mean ± SEM; Student’s t test was use for each phosphate-buffered saline (PBS) versus CBE comparison, ****P < 0.0001; ***P < 0.001. Each plot shown represents an individual mouse (pink, female; blue, male).