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The MS/MS spectra for the low-intensity peaks in the MALDI imaging spectra were acquired from the mouse brain extracted glycans to confirm their composition (Supporting Information S1).
> -wrap-foot> The same procedure was applied to 12 676 microarray gene expression profiles for the mouse brain, extracted from the GNF expression atlas (Su et al., 2004; see Supplementary Methods and Table S1).
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To determine whether the endogenous Tmub1/HOPS is localized in the membranous compartment, we fractionated mouse brain extracts into cytosolic and membranous fractions (Figure 2F).
The anti-pS34 antibody recognized AATYK1 immunoprecipitated from a mouse brain extract at a molecular weight ∼200 KDa (Fig. 5A, lane 1).
The possibility that lipin-1α stimulates the transcriptional activity of MEF2 by a direct coactivator mechanism is supported by our finding that lipin-1α coimmunoprecipitated with endogenous MEF2C in mouse brain extract (Fig. S4A) and with ectopically expressed MEF2C in HEK293A cells (Fig. S4B).
Mouse brain extract in SDS/PAGE loading buffer was obtained from Sigma Aldrich.
A higher protein concentration (40 μg/μL) of WT mouse brain extract decreased the adhesion of cells by approximately 75%.
(Bottom) GST pulldowns from a mouse brain extract using GST alone or GST fusions of OCRL as baits.
Both in the yeast extracts and in the mouse brain extract from Sigma, mouse RPS3A migrated approximately 6 kDa higher than its predicted molecular mass of 29.89 kDa.
Antibodies we generated against the mouse FLJ46154 protein recognize a protein of the predicted size for FLJ46154 in mouse brain extract (supplementary figure 6).
A more expansive huntingtin (htt) interactome subnetwork, comprising over 700 candidate proteins, was likewise identified in mouse brain extracts using AP-MS by Shirasaki et al. [ 58].
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