Fig 2: E1577K shows impaired Rac1-GEF activity. (A) Western Blot of anti-FLAG pulldown of Kalirin-7 and E1577K from HEK293T, with co-immunoprecipitation of transfected NR2B (Mock = non-transfected). (B) HEK293T cells were transfected with the indicated conditions and active-Rac1 was pulled down from lysates with PAK-PBD coated Sepharose beads. Positive (+ ve) and Negative (-ve) samples indicate Mock transfected lysates Loaded with GTP-?-S and GDP, respectively. Blots were probed with anti-FLAG and anti-Rac1. (C) Quantification of Active-RAC1 pulldown under the indicated conditions. *p < 0.05.

Fig 3: E1577K impairs surface expression of NR2B. (A) Representative regions of secondary dendrites with GFP-cell fill and NR2B surface stain. Insets show a single spine from each condition. Scale bar -10 µm. (B) Number of NR2B surface puncta normalized to total dendrite (GFP) area measured. (C) Average surface NR2B puncta size. (D) Average intensity of NR2B surface puncta. N = 2, 5 cells per replicate. (E) Regions of dendrites stained with Kalirin-7 and total NR2B to assess colocalization following overexpression of indicated constructs. Deconvoluted insets indicate Kalirin-7 and NR2B puncta adjacent within indicated spines. *p < 0.05. **p < 0.01. ***p < 0.005.