For the phosphopeptide competition assay, 2 mg of brain lysate from FLAG-LRRK2 Wt transgenic mice was incubated with different concentration of non-phosphopeptide or phophopeptide (S935) for 1 hour.
In their first paper of 2012, the authors injected recombinant human α-syn preformed fibrils or brain lysate from symptomatic transgenic mice overexpressing A53T α-syn [M83 mice, 22] into the cortex and striatum of asymptomatic transgenic mice.
Immunoblots of brain lysates from mutant animals showed a significant reduction in total Nrp1 protein (Fig. 1F).
Of note, SEC24B protein levels were increased in brain lysates from Sec24a gt/gt mice, with a potential slight increase in Sec24a +/gt heterozygous mice, though no differences were observed for SEC24C or SEC24D.
However, this signal was only observed with one of our two validated anti-G72 antibodies and a signal at the same apparent molecular weight was also detected in brain lysates from rat, which does not have a G72 gene, thus making it highly unlikely that this band represents G72.
(A ) Immunoblots of whole brain lysates from wild type, heterozygous, and knockout mouse at E18.5 show substantial reduction of synaptic proteins in knockout brain.
We therefore carried out an immunoblot analysis of brain lysates from 4-month-old Sp+/+, Sp Δ/+ and Sp Δ/Δ mice using the S51 antibody, directed against the N-terminal region of spastin (from amino acids 87 to 354) (Errico et al., 2004), which could thereby bind to the truncated protein.
(a) Immunoblot of lysates from brain lysates from wild-type (WT) and LAMP-2-deficient (LAMP-2-/y) mice as well as HeLa cells overexpressing eGFP, murine LAMP-1 (L1), murine LAMP-2A (L2A), murine LAMP-2B (L2B) or murine LAMP-2C (L2C).
Consistent with a role of PS in the proteolysis of neurexins, we found a significant accumulation of neurexin-CTFs in brain lysates from PS1−/− embryos and PS cDKO adult mice compared with control littermate animals (Fig. 4A).
