Figure 3.
Blocking endocytosis leads to increased CaV1.2-HA cluster density after 30 min. A, Experimental workflow. pf, Paraformaldehyde. B, Dynasore blocks dynamin-dependent endocytosis and membrane recycling. C, D, Live cell staining of neurons cotransfected with CaV1.2-HA and eGFP shows that blocking endocytosis with 80 μm Dynasore (Dyn) for 15 and 30 min does not alter the labeling pattern of CaV1.2-HA clusters in dendritic shafts and spines. Scale bar, 5 μm. E, F, Quantification of CaV1.2-HA clusters indicates no significant change in clusters density and size after 15 min of Dynasore treatment, but a significant 26.6 ± 12.2% and 33.1 ± 11.8% increase of cluster density on shafts and spines after 30 min of Dynasore treatment (mean ± SEM; N = 23–31 analyzed dendritic segments in 3–4 separate experiments; *p = 0.02–0.05, significance, t test).