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Figure S1. Association of stonin 2 with synaptotagmin 1. (A) 6 μg GST–synaptotagmin 1 cytoplasmic domain (C2A, C2B, or C2AB) fusion proteins (bottom) were analyzed in pulldown experiments for their ability to associate with HA–stonin 2^WT from HEK293 cell lysates (Materials and methods). Samples were analyzed by immunoblotting (top) for HA–stonin 2, AP-2μ, or clathrin heavy chain (CHC). 8% standard (STD), 8% of the total input (cell extract). (B) Same as in A, but with an AP-2–binding–deficient HA–stonin 2^δWFδNPF mutant. (C) GST-C2B wild-type or mutant proteins (Fig. 2 B) were analyzed in pulldown experiments for their ability to associate with HA–stonin 2^δWFδNPF from HEK293 lysates. Samples were analyzed by immunoblotting for HA–stonin 2, AP-2β, and clathrin heavy chain. 3% of the input was loaded as standard.