P27 25614 1 ap

RIPA lysis buffer (Thermo Scientific) containing phosphatase and protease inhibitors was used to extract cellular proteins, and the BCA Protein Assay Kit (Thermo Fisher Scientific) was used to measure proteins, followed by protein blotting. Information on all primary antibodies used in this study is as follows: TUBA1C (ab222849, Abcam), β-Actin (#3700, Cell Signaling Technology), Bcl-2 (#15,071, Cell Signaling Technology), Bax (#41,162, Cell Signaling Technology), Cyclin B1 (#12,231, Cell Signaling Technology), CDK1 (10762-1-AP, Proteintech), P27 (25614-1-AP, Proteintech) and P21 (10355-1-AP, Proteintech).

Western blot was performed according to a previous publication (Zhang et al. 2017 (link)). The antibodies and the dilution used in the detection were as follows: GSG2 (NBP1-26,626, 1:3000 dilution, NOVUS, USA), GAPDH (10,494-1-AP, 1:5000 dilution, Proteintech Group, China), GSK-3α/β (5676, 1:1000 dilution, CST, MA, USA), phospho-GSK-3α(Ser21) (8452, 1:1000 dilution, CST, MA, USA), phospho -GSK-3β(Ser9) (5558, 1:1000 dilution, CST, MA, USA), p27(25,614-1-AP, 1:1000 dilution, Proteintech Group, China), p21(10,355-1-AP, 1:1000 dilution, Proteintech Group, China), cdc2(9116,1:1000dilution, CST, MA, USA), phospho-cdc2 (Tyr15) (4539,1:1000dilution, CST, MA, USA), cyclin A(4656,1:1000dilution, CST, MA, USA), cyclin B(12,231,1:1000dilution, CST, MA, USA). All the protein bands were exposed with Super ECL Detection Reagent (36,208, YEASON, China) on the Chemiluminescence imaging system (Cytiva, USA). The protein bands were analyzed by Image J software (NIH, USA). All the blots were cut prior to hybridisation with antibodies, so some full-length blots cannot be provided. However, all membranes are cut along the corresponding upper and lower markers according to the molecular weight of proteins to ensure the accuracy of the experiment. All the original images are displayed in the supplementary material.