Bcl 2

The cortex tissue of the left brain after 24 hours post-HI or cells treated in each group were centrifuged (1000 ×g, 15 min) to obtain the supernatants. After analyzing the concentration of protein, the protein sample was electrophoresed on SDS-PAGE and then transferred to the membrane. After blocking, add PIK3R2 (1:500, orb76194, Biorbyt, Cambridge, UK), LC3B (1:1,000; 2775S, Cell Signaling Technology, Danvers, MA, USA), Beclin1 (1:1,000; 3738s, Cell Signaling Technology, Danvers, MA, USA), p62 (1:500, orb89844), Cleaved-capase-3 (1:500, orb106556), Bax (1:500, orb224426), Bcl-2 (1:500, orb228150), β-actin (1:2000, orb178392) for incubation. p62, Cleaved-capase-3, Bax, Bcl-2 and β-actin are all from Biorbyt. Finally, it was incubated with goat anti-rabbit IgG antibody (1:1,000; ABIN101988) for 60 minutes. For 3-5 minutes, the membranes were seen using an ECL luminous substrate. Using ImagePro Plus, proteins were scanned, measured, and normalized to β-actin.

Shells of pearl oysters (Pinctada maxima) were collected from Iki Bay (Nagasaki, Japan). Antibodies against sirtuin 1, BDNF, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cyclin-dependent kinase inhibitor 2A (p16) were purchased from Biorbyt (San Francisco, CA, USA).