CRACR2a cortical puncta recruit dynein to detach from the actin cortex and travel to the MTOC. (A) Colocalization between CRACR2a cortical puncta and dynein (visualized by mCherry-Tctex3, a dynein light chain). Scale bar: 5 µm. Inset scale bar: 2 µm. (B) Imaging CRACR2a cortical puncta dynamics on actin (visualized by mCherry-F-tractin) and microtubules (visualized by SiR-Tubulin) using a spinning disk confocal microscope. Kymograph analysis along the indicated line is shown on the right. In total, 28 cells were imaged continuously for 50 s each, and 16 events of a cortical punctum moving on a microtubule were observed. Scale bar: 5 µm. (C) Changes in CRACR2a cortical puncta dynamics in response to cytoskeleton disassembly by nocodazole (5 µM) and/or latrunculin A (2 µM). The cells are treated as indicated by the diagrams shown on the left side. Scale bar: 5 µm.