Fig. 8
RIM1 acts as an effector of Rab11 and participates in the recycling of NMDARs. a–c Rab11 is involved in the RIM1/PSD-95/NMDAR complex in mouse cortex. Extracts of mouse cortex were immunoprecipitated with antibody against PSD-95, Rab11, or RIM1, and blotted with the corresponding antibodies. d GST-RIM11-399 interacts directly with Rab3 and Rab11, but not with Rab5, in the presence of GTP-γS. e GST-RIM11-55 has a direct interaction with Rab3, but not with Rab5 or Rab11, in the presence of GTP-γS. f RIM1 KD significantly increases the colocalization of Rab11 and GluN2B. Left: cultured hippocampal neurons were transfected with control RNAi (n = 23 neurons from three independent cultures) or RIM1 RNAi (n = 32 neurons from three independent cultures) plasmids at DIV6 and were then co-immunostained with antibodies against GluN2B and Rab11 at DIV10 and imaged by 3D-SIM. The lower four lines (“one slice”) show one of the Z slices. Scale bar, 3 µm. Right: statistical analysis of colocalization of GluN2B and Rab11; t-test, *p < 0.05. g RIM1 KD significantly decreases the colocalization of SNAP25 and GluN2B. Left panel: cultured hippocampal neurons were transfected with control RNAi (n = 15 neurons from three independent cultures) or RIM1 RNAi (n = 12 neurons from three independent cultures) plasmids at DIV6. The neurons were immunostained with antibodies against GluN2B and SNAP25 at DIV10 and imaged by 3D-SIM. The lower four lines (“one slice”) show one of the Z slices. Scale bar, 3 µm. Right panel: statistical analysis of colocalization of GluN2B and SNAP25; t-test, *p < 0.05
