Western Blot Analysis of Brain Proteins

Mice were decapitated at the time of each experimental procedure, and the right cerebral hemisphere was separated and homogenized in RIPA buffer (Servicebio, Wuhan, China). Western blotting was performed as previously described [20 (link)]. The primary antibodies included anti-MST4 (1 : 1000, Proteintech), anti-AKT (1 : 1000, Cell Signaling Technology), anti-pAKT (1 : 1000, Cell Signaling Technology), anti-LC3A/B (1 : 1000, Cell Signaling Technology), and β-actin (1 : 1000, Cell Signaling Technology).

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Wu X., Wu J., Hu W., Wang Q., Liu H., Chu Z., Lv K, & Xu Y. (2020). MST4 Kinase Inhibitor Hesperadin Attenuates Autophagy and Behavioral Disorder via the MST4/AKT Pathway in Intracerebral Hemorrhage Mice. Behavioural Neurology, 2020, 2476861.

Publication 2020

RIPA buffer anti-AKT anti-pAKT anti-LC3A/B β-actin

Actin Antibodies Buffer Mice Right cerebral hemisphere Ripa

Corresponding Organization : First Affiliated Hospital of Wannan Medical College

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Variable analysis

independent variables

Experimental procedure (time of decapitation)

dependent variables

Expression levels of MST4, AKT, pAKT, and LC3A/B proteins

control variables

Right cerebral hemisphere

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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