Figure 2: Impact of gatastatin on MT regrowth in HeLa cells.

(a) The impact of gatastatin on MT nucleation by centrosomes in interphase. After nocodazole and ice treatments, MT nucleation (1 min at 37 °C) in the presence of 30 μM of gatastatin or the DMSO solvent control was monitored. Red, green and blue in the merged image represent EB1, α-tubulin and pericentrin, respectively. Scale bar, 10 μm. Three independent experiments were performed and one representative experiment is shown. (b) The area of EB1 around centrosomes in a was calculated using the ImageJ software from 97 (control) or 84 (gatastatin) centrosomes. Error bars represent s.e.m. Two-tailed, unpaired Student’s t-test was used to obtain P value. (c) The impact of gatastatin on MT nucleation by centrosomes in mitosis. After STLC and ice treatments, spindle MT nucleation (1 min at 20 °C) in the presence of 30 μM of gatastatin or the DMSO solvent control was monitored. Red, green and blue in merged image represent EB1, α-tubulin and pericentrin, respectively. Scale bar, 10 μm. Three independent experiments were performed and one representative experiment is shown. (d) The area of EB1 around centrosomes in c was calculated using ImageJ software from 22 (control) or 17 (gatastatin) monopolar spindles. Error bars represent s.e.m. Two-tailed, unpaired Student’s t-test was used to obtain P value.