Fig 1: GSK3 paralogs localise to distinct locations in mouse and human platelets. Platelets and MEFs were stained with antibodies specific for GSK3α (D80D1, #4818 CST) and GSK3β (MRU PPU) and imaged using a Leica TCS SP8 STED system: (A) Representative images of WT, GSK3α KO and GSK3β KO MEFs stained with antibodies against GSK3α (yellow) and GSK3β (magenta) in the presence of 4′,6-diamidino-2-phenylindole (DAPI, blue) indicated that antibodies used for localisation analyses are specific for GSK3α and GSK3β. Bar represents 6 µm. (B) Immunoblotting of WT, GSK3α KO, and GSK3β KO MEFs for GSK3α, GSK3β, and pGSK3α/βS21/9 verifies that MEFs have either GSK3α or GSK3β deleted. Data are representative of 3 independent experiments. (C) Representative images of mouse platelets showing GSK3α (yellow) and GSK3β (magenta). Bar represents 1 µm. (D) Representative images of human platelets showing GSK3α (yellow) and GSK3β (magenta). Bar represents 1 µm. (E) Pearson’s correlation coefficient analysis demonstrates that GSK3α and GSK3β are unlikely to interact physically in mouse (mGSK3α vs. mGSK3β, 62 images) and human (hGSK3α vs. hGSK3β, 22 images) platelets.