Movie 5.

Representative single plane spinning disk microscopy time lapse demonstrating non-responsiveness of Rab7 endosomes to acute TRPML1 agonism. Stable EmRab7WT NRK cells were treated with the TRPML1 agonist ML-SA1 to stimulate Ca²⁺ release from lysosomes in the absence of membrane disruption. Time lapse was captured for 20 minutes at 3 sec per frame. 20μM ML-SA1 was added directly to stage at 10 minutes. Scale bar = 10μm, frame rate = 20fps.