Expression of SnxA fusions in mammalian cells. (A–F) Representative images and quantification of either full-length SnxA or 2xPX GFP fusions expressed in either HEK293, MIA PaCa-2, or RPE-1 cells. Top panels of A, C, and E are maximum intensity projections of a spinning-disc Z-stack, with single sections and enlargements of the boxed regions below. Cells were either left untreated (−) or pretreated with 1 μM apilimod (Ap; +) for 1 h prior to imaging. Arrowheads indicate the GFP-positive structures that were quantified over three independent experiments in B, D, and F. 30 cells per experiment for three independent experiments were scored for each condition and P values were calculated by unpaired T test (n = 3, *P < 0.05, **P < 0.01); scale bars represent 10 μm. (G) Shows an image from a timeseries (Video 1) of a HEK293 cell expressing GFP-SnxA just after addition of Texas-Red dextran. Scale bars represent 2 μm.