Fig 1: Ketamine enhances PMCA2b interaction with NMDAR1. (A) The expression of Grin1 encoding NMDAR1 subunit evaluated with real-time PCR and calculated using the 2-??Ct method. The expression level in GFP control was taken as 1, n = 4. (B) NMDAR1 protein level evaluated with the Western blot. GAPDH was taken as a loading control. The representative blots are presented. (C) Quantification of NMDAR1 protein level following normalization to endogenous Gapdh level, n = 3. AU, arbitrary units. (D) The mRNA and (E) protein levels of NMDAR2a. (F) Quantification of NMDAR2a protein level following normalization to endogenous Gapdh level, n = 3. (G) The mRNA and (H) protein levels of NMDAR2b. (I) Quantification of NMDAR2b protein level following normalization to endogenous Gapdh level, n = 3. (J) Co-immunoprecipitation of NMDAR1 and PMCA2b. Lysates from differentiating H19-7 cells were used to measure NMDAR1 protein level (input) in saline- and ketamine-treated conditions. Negative controls also included sepharose-linked secondary antibodies (IgG) and sepharose beads only. Representative blots are presented. (K) Quantitative densitometric analysis of band intensity in PMCA2b-expressing cells. The results are presented as arbitrary units defined as the optical density per mg protein, n = 3. *P < 0.05, **P < 0.01.