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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: X-linked intellectual disability gene CASK regulates postnatal brain growth in a non-cell autonomous manner

Fig. 5

CASK is present in cytosol as a part of large protein complexes. a Western blot showing endogenous level of CASK, synaptophysin, ATP synthase β subunit and tubulin from CASK (+/+) and CASK (+/-) mutant mice (n = 3). b Western blot quantitation in CASK (+/-) mice relative to CASK (+/+) mice. Data is normalized to tubulin and expressed relative to wild-type levels. Bar graphs are plotted as mean ± SEM. (* indicates p < 0.05; n = 3). c Wild-type mice brains were initially fractionated to separate synaptosomal and cytosolic fractions. The synaptosomal fraction was solubilized initially in Triton X-100 and then in deoxycholic acid (DCA). Samples were blotted for the indicated antigen. d Wild-type mouse brain homogenate was solubilized in Triton X-114 at 4 °C, samples were warmed up to 37 °C and centrifuged to separate into an aqueous phase, a detergent phase and a pellet. Equal dilutions of each phase were blotted for the indicated proteins. e Aqueous phase (after Triton X-114 phase separation) from brain containing CASK was carefully layered on a 10-40 % continuous glycerol gradient. Following ultracentrifugation, samples were blotted for CASK and liprin-α3. Fig. also shows pure recombinant CASK. Arrows indicate the fractions where protein standards of 150 kDa and 460 kDa were observed. f GST-pulldown assay performed from rat brain homogenates using either full-length GST-CASK or GST (negative control) immobilized on glutathione resin. Synaptophysin (negative control); liprin α2 (positive control); ATPsynβ (ATP synthase β subunit); and IDH (isocitrate dehydrogenase)

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