Took a 2 g sample, added liquid extraction buffer and protease inhibitor in a certain ratio after grinding in liquid nitrogen in a frozen state, vortexed for 10 min. Added the same volume of Tris-saturated phenol (pH 8.0) and vortexed for 10 min; centrifuged at 12000 g at 4 °C for 20 min to obtain a phenol phase; took the phenol phase into a new centrifuge tube, added an equal volume of extraction buffer, vortexed Shake for 10 min; centrifuged at 12,000 g for 20 min at 4 °C to obtain a phenol phase; took the phenol phase into a new centrifuge tube, added the pre-chilled ammonium acetate methanol solution in proportion, and precipitated the protein overnight at − 20 °C; centrifuged at 12000 g for 20 min at 4 °C; discarded Supernatant, added 90% acetone and vortexed to mix and washed twice. Suspended the pellet with an appropriate volume of lysate (8 M urea + 1% SDS, protease inhibitor has been added) to fully dissolved the sample protein. Centrifuged at 4 °C, 12000 g for 20 min. Collected the supernatant. The quantitative results of BCA were shown in Fig. S2 and Table S1.

Lysis buffer (1% SDS, 8 M urea, 1x Protease Inhibitor Cocktail (Roche Ltd. Basel, Switzerland) was added into the samples. The lysis was performed by sonication on ice for 2 min and kept on ice for 30 min. After centrifugation at 15000 rpm for 15 min at 4 °C, the supernatant was collected and transferred to a new Eppendorf tube.

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