Supplementary Figure 4: GluN1 N616R pore mutation renders NMDARs Ca2+-impermeable.

a) Representative I-V traces from GluN1^WT and GluN1^N616R transfected N2a cells in normal (2mM) and high (40mM) extracellular Ca²⁺. Cells were voltage-clamped in whole-cell configuration (V_hold = -60mV), and voltage ramps were applied from -80mV →; +80mV in the presence of NMDA(100μM)/Gly(10μM). Ca²⁺ permeability was assessed by increasing bath concentrations from 2mM to 40mM. Insets show expression of GluN1^WT- and GluN1^N616R–mCerulean+ constructs. b) Quantification of average shift in reversal potential in different [Ca²⁺]_o.