Adaptor interaction sites on the CHC NTD directly interact with GTSE1. (A) Immunoblots of cell lysates (input) and IPs of GTSE1 and CHC in U2OS and GTSE1 knockout (GTSE1^KO) cells. Immunoblots with anti-GTSE1 or anti-CHC antibody. (B) Immunoblot of cell lysates (input) and IPs of GTSE1-GFP in mESCs. Immunoblots with anti-CHC (top) and anti-CHC and anti-GFP (bottom) antibodies. (C) GST-CHC pulldown of purified recombinant GTSE1. Immunoblot with anti-GTSE1 antibody. Coomassie blue of GST inputs is presented. (D) Pulldown analysis of the interaction between purified GTSE1-GFP and CHC fragments (numbers designate amino acids) fused to GST. Immunoblot with anti-GFP antibody. Coomassie blue of GST inputs is presented. (E) Pulldown analysis of the interaction between purified GTSE1 and CHC(1–330) fused to GST. CHC was mutated at site 1 (S1M: T87A, Q89A, K96E, K98E), site 2 (S2M: Q152L, I154Q), or site 3 (S3M: R188A, Q192A). Immunoblot with anti-GTSE1. Coomassie blue of the GST inputs is presented.