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. 2019 Dec 9;30(5):3030–3043. doi: 10.1093/cercor/bhz292

Figure 1.

Figure 1

SEPT7 interacts with KIF20A. (A and B) Reciprocal coimmunoprecipitation (co-IP) of GFP-SEPT7 with Flag-KIF20A. Expression plasmids were cotransfected into Hek293 cells. Coexpressed proteins in cell lysates were precipitated for KIF20A (anti-Flag) or SEPT7 (anti-GFP) followed by western blot for the other tagged protein. (C) Two deletions of KIF20A were made and fused with Flag tag (Flag-D1 and Flag-D2). Expression plasmids of Flag-KIF20A or Flag-deletions were cotransfected with GFP-SEPT7 into Hek293 cells. In the top panel, coexpressed proteins were precipitated for KIF20A or deletions (anti-Flag) followed by western blot for GFP-SEPT7 (anti-GFP). In the bottom control panel, the above anti-Flag co-IPs were blotted by anti-Flag antibody to confirm IP of the Flag-tagged deletions. (D) Co-IP of endogenous SEPT7 and KIF20A proteins. Cell lysates were made from the E15.5 mouse brains and proteins were immunoprecipitated with anti-KIF20A antibody followed by western blot for SEPT7. (E) In dissociated dividing cortical NPCs, PLA using both SEPT7 and KIF20A antibodies could detect specific PLA events in discrete areas. Arrows in high magnification images indicate the PLA event detected in the ICB of a dividing NPC at telophase. (F) No PLA events were detected in cultured NPCs when only one antibody (KIF20A antibody, green) was present.