Figure 4.
Clusters of CaV1.2-HA channels do not colocalize with Tf-488-labeled recycling endosomes. A, C, Experimental workflow. pf, Paraformaldehyde. B, Hippocampal neurons incubated for 30 min with Tf-488 and subsequently labeled with phalloidin-Alexa Fluor 594 to outline dendritic spines shows abundant uptake of Tf-488 into endocytic compartments in the dendritic shaft and, to a lesser degree, in the spines. D, Simultaneous incubation with Tf-488 and anti-HA and subsequent immunolabeling of permeabilized neurons demonstrates little to no colocalization of CaV1.2-HA and Tf-488 into endocytic compartments after 30 min. E, If CaV1.2-HA (red) would be subject to rapid recycling, co-uptake with Tf-488 (green) would have been expected. Scale bars, 5 μm.