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. Author manuscript; available in PMC: 2023 May 1.
Published in final edited form as: Nat Chem Biol. 2022 Aug 22;18(11):1224–1235. doi: 10.1038/s41589-022-01096-2

Figure 4: Lattice spacing governs MAP cooperativity in vivo.

Figure 4:

a. Time lapse micrographs of U-2 OS cells expressing eGFP-tau treated with 0.01 μM taxol (top panels) or DMSO (bottom panels). Zoom-ins of the indicated region are provided in the top left corner of the original micrograph. Scale bars: 10 μm. b. Coefficient of variation of eGFP-tau cells 10 minutes after taxol or DMSO treatment, normalized to the coefficient of variation before treatment (at t=−1 min). Coefficient of variation was calculated over the whole cell (eGFP-tau after taxol treatment: 66.6 ± 8.0 (mean ± s.d.); n = 18 cells, 5 experiments; eGFP-tau after DMSO treatment: 96.8 ± 11.8 (mean ± s.d.); 17 cells, 7 experiments, two-sided t-test, p<0.0001). c. Time lapse micrographs of U-2 OS cells expressing eGFP-MAP4 treated with 0.01 μM taxol (top panels) or DMSO (control, bottom panels). Zoom-ins of the indicated regions are provided in the top left corner of the original micrograph. Scale bars: 10 μm. d. Coefficient of variation of eGFP-MAP4 cells 10 minutes after taxol or DMSO treatment, normalized to the coefficient of variation before the same treatment (at t=−1 min). The coefficient of variation was calculated over the whole cell (eGFP-MAP4 after taxol treatment: 93.8 ± 20.4 (mean ± s.d.); n = 23 cells, 5 experiments; eGFP-MAP4 after DMSO treatment: 92.7 ± 7.5 (mean ± s.d.); n=23 cells, 5 experiments, two-sided t-test, p=0.7936). e. Zoom in from movie as shown in a showing the progression of tau signal on a single microtubule (yellow arrowhead) initially fully covered by eGFP-tau after taxol treatment. Sketches of the micrographs (panels next to the original micrographs) indicate the size and locations of fissures in tau signal that appear after taxol treatment. Scale bar: 2 μm. f. Fluorescence kymograph of the microtubule presented in e showing the fissures (yellow arrows) appearing in the eGFP-tau signal and the disassembly of the tau envelopes from their boundaries. Scale bars: horizontal 1 μm, vertical 10s. g. Fluorescence kymograph of tau envelope disassembly in vitro showing striking resemblance with the in vivo observations in f. Scale bars: horizontal 2 μm, vertical 10s.