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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Mol Cell Neurosci. 2013 Jul 31;56:10.1016/j.mcn.2013.07.010. doi: 10.1016/j.mcn.2013.07.010

Figure 5. PICK1 and Arc knockdown prevent increased AMPAR cycling of RGCs in the absence of activity.

Figure 5

A. Images depict expression of PICK1 or Arc following siRNA treatment (scale bar=10μm). B. Surface GluA2 levels were measured immunocytochemically in cultured RGCs 72 hours after transfection with PICK1 or Arc siRNA and compared to levels in control siRNA transfected cells. Knockdown of Arc but not PICK1 increased surface GluA2 levels (n>5, * p≤ 0.05). C–F. Cultured RGCs were transfected with siRNA against PICK1 or Arc. After 72 hours constitutive AMPAR endocytosis was monitored immunocytochemically in cultures maintained in the absence of synaptic activity (−Activity) a condition under which ARC and PICK1 are normally elevated. Effects of PICK1and Arc siRNA were also measured in the presence of activity (+ Activity). C,E. Images depict dendritic labeling of surface GluA2 in PICK1(C) or Arc(E) siRNA transfected RGCs at 0 and 45 minutes after initial immunolabeling (scale bar=5μm). D,F. Quantitation of surface GluA2 fluorescence in experiments as in B and D demonstrates that knockdown of PICK1 (n>5, *p<0.05) ( D) and Arc (n≥5, **p<0.01) (F) both prevent the increased constitutive GluA2 trafficking normally observed in RGCs in the absence or presence of synaptic activity (−Activity or +Activity) (see Fig 4B).