Fig. 6

Parkin deficiency impairs GluN1 internalization and recycling. a Representative images of 14–16 DIV hippocampal neurons expressing soluble GFP control/shParkin/shParkin-WT constructs, immunostained for cell surface (surf) and internalized (inter) GluN1 after 0 and 15 min internalization at 37 °C. Scale bars, 10 μm. b Representative images of 14–16 DIV hippocampal neurons expressing the same constructs as in (a), immunostained for cell surface (surf) and recycled (rec’d) GluN1 after 30 min internalization followed by 0 and 60 min recycling at 37 °C. Scale bars, 10 μm. c Quantification of GluN1 internalization at 15 min, expressed as the ratio of internalized to cell surface GluN1 and normalized to GFP control condition (n ≥ 50 fields of view per condition with > 50 GluN1 puncta per field, results confirmed in 3 independent experiments; ***P < 0.001, unpaired t test, error bars represent SEM). d Quantification of GluN1 recycling at 60 min, expressed as the ratio of recycled to internalized GluN1 and normalized to GFP control condition (n ≥ 55 fields of view per condition with > 50 GluN1 puncta per field, results confirmed in 3 independent experiments; ***P < 0.001, unpaired t test, error bars represent SEM)