Anti mbp

Vectors for all the following analyses were constructed by introducing the full-length coding sequences of the genes into empty vectors using the in-fusion strategy. Also, see our previous studies for the available vectors related to GSK2 and aGSK2 (Liu et al., 2021 (link); Tong et al., 2012 (link)). All primers and resulting vectors are listed in Supplemental Table 1. A yeast two-hybrid assay was performed using the Matchmaker Gold Yeast Two-Hybrid System (Clontech) according to the product instructions. The pull-down assay and expression and purification of fusion proteins were performed as described previously (Liu et al., 2022 (link)). BiFC, luciferase complementation, and Co-IP assays were performed as detailed previously (Chen et al., 2008 (link); Xiao et al., 2020 (link); Liu et al., 2022 (link)). The fluorescence was detected with a confocal fluorescence microscope (LSM 980, ZEISS). The luminance signal was detected with an imaging system equipped with a cold charge-coupled device camera (LB 985, NightSHADE with indiGO software). Detection of proteins by immunoblotting was performed using tag antibodies including anti-FLAG (1:1000, Sigma), anti-GFP (1:1000, Abmart), anti-GST (1:2000, Abmart), and anti-MBP (1:2000, Abmart).