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. Author manuscript; available in PMC: 2014 Nov 25.
Published in final edited form as: Dev Cell. 2013 Nov 7;27(4):425–437. doi: 10.1016/j.devcel.2013.10.007

Figure 6. The VPS41 Clathrin Heavy Chain Repeat Is Required for Regulated Secretion.

Figure 6

(A) Model of VPS41 showing the putative AP-3 binding site at the N terminus as well as the CHCR at the C terminus. The ΔC mutant contains residues 1–595 and ΔN36 residues 37–853.

(B) PC12 cells were cotransfected with control or VPS41 siRNAs, ANF-GFP, and RNAi-resistant VPS41 full-length (WT) and C-terminal deletion (ΔC) rescue constructs where indicated, and ANF-GFP secretion was measured as described in Figure 1E. *p < 0.05 relative to stimulated secretion from control (n = 3–4). Bar graphs represent mean ± SEM.

(C) Western analysis of the HA-tagged, RNAi-resistant constructs transfected in duplicate into PC12 cells shows equivalent expression of WT and ΔC VPS41.

(D) PC12 cells were cotransfected with control or VPS41 siRNAs, ANF-GFP, and RNAi-resistant WT or ΔN36 VPS41, and ANF-GFP secretion measured as described in Figure 1E. *p < 0.05 by ANOVA followed by Tukey posthoc testing, relative to stimulated secretion from control (n = 4). Bar graphs represent mean ± SEM.