Figure 4.

Assays for AP-1 Function

(A and B) Cells coexpressing γ-FKBP and Mito-YFP-FRB were depleted of endogenous γ with siRNA, and incubated with anti-CIMPR for 45 min at 37°C, either without (A) or with (B) rapamycin. See also Figure S3. Scale bar, 20 μm.

(C) Cells coexpressing γ-FKBP and Mito-YFP-FRB were depleted of endogenous γ with siRNA. Half of the cells were treated with rapamycin for 10 min before homogenization, then CCVs were isolated and western blots probed with the indicated antibodies.

(D) Blots of whole-cell homogenates and CCV preparations from a conventional AP-1 γ knockdown, probed with the same antibodies as in (C).