Fig. 1. Nova-1 interacts with 5-HT6R in vitro and in vivo. (A) Schematic diagrams showing the structure of 5-HT6R, the CT (carboxyl-terminus) of 5-HT6R as bait and complete Nova-1 protein as prey. (B) When the AH109 strain with 5-HT6R CT and the Y187 yeast strain with Nova-1 were mated, a blue color was detected. (C) GST pull-down assay showed that Flag-Nova-1 specifically interacted with GST-fused CT of 5-HT6R (GST-6RCT). (D) Full-length Myc-tagged 5-HT6R (Myc-5-HT6R) signal was detected in the immunoprecipitation with anti-Flag. (E) Flag-Nova-1 signal was detected in the immunoprecipitation with anti-Myc. (F) Western blot with anti-5-HT6R or anti-Nova-1 antibody showed the expression patterns of 5-HT6R and Nova-1 in the rat brain. (G) GST-6RCT interacted with endogenous Nova-1 in the rat brain. (H) Co-immunoprecipitation assay confirmed the interaction between endogenous 5-HT6R and Nova-1 in the rat brain. (I) When GST pull-down assays were performed between Nova-1 and CTs of several serotonin receptors (GST-4RCT, GST-6RCT and GST-7BRCT), Nova-1 only interacted with 6RCT but not 4RCT and 7BRCT. GAL4, galactose-responsive transcription factor GAL4; BD, binding domain; AD, activation domain; C, Cortex; Hi, Hippocampus; S, Striatum; B, Brainstem; Hy, Hypothalamus.
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