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. 2024 Feb 22;13:e91719. doi: 10.7554/eLife.91719

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© 2024, Gonzalez et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 2. — (A) Left: Simulations were performed in which the EB1 protofilament-edge on-rate was set to zero, and the closed-lattice on-rate was gradually increased. Right: Simulated kymographs in which the EB1 protofilament-edge on-rate was set to zero, and the on-rate at closed-lattice sites was gradually increased (scale bars: 2 µm and 10 s). (B) Left: Simulations were performed in which the closed-lattice on-rate remained constant at its baseline (non-zero) value, and the protofilament-edge on-rate was gradually increased. Right: Simulated kymographs in which the closed-lattice on-rate remained constant at its baseline (non-zero) value, and the protofilament-edge on-rate was gradually increased (scale bars: 2 µm and 10 s). (C) Left: Simulated images of EB1-GFP tip tracking over a range of closed-lattice on-rates (scale bar: 1 µm). Center: Line scans from simulated images of EB1-GFP intensity for a range of closed-lattice on-rates (error bars, SEM). Right: Tip:Lattice EB1-GFP intensity ratio vs closed-lattice on-rates in the simulation (error bars, SEM). The tip:Lattice EB1-GFP intensity ratio decreases with increasing closed-lattice on-rates. (D) Simulated kymograph in which the closed-lattice on-rate is set to zero partway through the simulation, and later returned to its baseline value (scale bars: 2 μm and 20 s). (E) Left: Simulated images of EB1-GFP tip tracking over a range of protofilament-edge on-rates (scale bar: 1 µm). Center: Line scans from simulated images of EB1-GFP intensity for a range of protofilament-edge on-rates (error bars, SEM). Right: Tip:Lattice EB1-GFP intensity ratio vs protofilament-edge on-rates in the simulation (error bars, SEM). Localization to the microtubule tip increases with increasing protofilament-edge on-rates. (F) Simulated kymograph in which the protofilament-edge on-rate is set to zero partway through the simulation and later returned to its baseline value (scale bars: 2 μm and 20 s). (G) Top: Representative images of EB1-GFP tip tracking for increasing Protofilament-edge:Closed-lattice on-rate ratios. Bottom: Tip:Lattice EB1-GFP intensity ratio for increasing Protofilament-edge:Closed-lattice on-rate ratios (GDP:GTP off-rate ratio is constant and set to 12:1). The experimentally measured Protofilament-edge:Closed-lattice on-rate ratio is 50–100 (Reid et al., 2019) (gray dashed boxes). (H) Top: Representative images of EB1-GFP tip tracking for increasing GDP:GTP closed-lattice off-rate ratios. Bottom: Tip:Lattice EB1-GFP intensity ratio for increasing GDP:GTP closed-lattice off-rate ratios (Protofilament-edge:Closed-lattice on-rate ratio is constant and set to 50:1). The experimentally measured GDP:GTP closed-lattice off-rate ratio is 6–12 (Maurer et al., 2011) (gray dashed boxes). (I) Top: Representative images of EB1-GFP tip tracking without protofilament-edge binding. Bottom: Tip:Lattice EB1-GFP intensity ratio for experimentally measured GDP:GTP closed-lattice off-rate ratios, with (red) or without (blue) EB1 binding at protofilament-edge sites.

Figure 2—source data 1. Data for Figure 2C, E, G, H, and I.

elife-91719-fig2-data1.xlsx^{(327.7KB, xlsx)}