The GFP-ML1Nx2 probe does not dissociate from the LAMP1-positive membranes in response to blockade of PtdIns(3,5)P₂ synthesis.

Images show a representative cell expressing the indicated constructs before and 3 h after treatment with 1 μM YM201636, which inhibits PIKfyve that synthesizes PtdIns(3,5)P₂. The montage images at the bottom are from the boxed region above, and are separated by 10 min intervals. The graph at right shows mean fluorescence intensity at LAMP1-positive membranes normalized to the whole cell for the indicated construct (data are means ± s.e.m. of 29 cells from three independent experiments). No dissociation of the GFP-ML1Nx2 is observed despite extensive vacuolation of the cells in response to YM201636 treatment. Scale bar = 15 μm.