Figure 8.
Figure 8. AP-3 RNAi affects the membrane composition of LDCVs. PC12 cells were transfected twice with 50 nM AP-3δ or control siRNA, and the postnuclear supernatant obtained 2–3 d after the second transfection separated by equilibrium sedimentation through 0.6–1.6 M sucrose. (A) Fractions were collected from the top of the gradient and assayed for VAMP2, VAMP3, and synaptotagmin 1 (syt1) by quantitative fluorescent immunoblotting. (B) The amounts in each fraction in light membranes (peak I) and LDCV fractions (peak II) are expressed as a percentage of total gradient immunoreactivity. AP-3 RNAi greatly reduces the LDCV peak of synaptotagmin 1 and shifts the synaptotagmin 1 immunoreactivity toward lighter fractions. *, P < 0.05 relative to control by two-tailed Student’s t test (n = 3 transfections). Error bars indicate SEM.

AP-3 RNAi affects the membrane composition of LDCVs. PC12 cells were transfected twice with 50 nM AP-3δ or control siRNA, and the postnuclear supernatant obtained 2–3 d after the second transfection separated by equilibrium sedimentation through 0.6–1.6 M sucrose. (A) Fractions were collected from the top of the gradient and assayed for VAMP2, VAMP3, and synaptotagmin 1 (syt1) by quantitative fluorescent immunoblotting. (B) The amounts in each fraction in light membranes (peak I) and LDCV fractions (peak II) are expressed as a percentage of total gradient immunoreactivity. AP-3 RNAi greatly reduces the LDCV peak of synaptotagmin 1 and shifts the synaptotagmin 1 immunoreactivity toward lighter fractions. *, P < 0.05 relative to control by two-tailed Student’s t test (n = 3 transfections). Error bars indicate SEM.

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