Figure 1.
MTs undergo a centrosomal to non-centrosomal transition during GBE. (A–C) MTs marked with Jupiter:GFP at level of centrosomes 20 min pre-GBE (A), at GBE onset (B), and 20 min into GBE (C). Arrows point to centrosomal MT. (A′–C′) Perinuclear MT pools (green) at 20 min pre-GBE (A′), at onset (B′), and 20 min into GBE (C′). (A′’–C′′) Orthogonal projection of MT and nuclei in epithelium 20 min before GBE (A′′), at onset (B′′), and 20 min into GBE (C’’). (A‴–C‴) Line scan intensity plot for MT as denoted in the lines (blue = 2 μm above nuclear midplane, red = at nuclear midplane) shown in A′′, B′′, and C′′, respectively. (D) Normalized intensity of centrosomal MTs at 0 and 20 min GBE; n = 150 centrosomes for each time point, k = 3 embryos. (E) Normalized intensity of perinuclear MTs at 0 and 20 min GBE; n = 80 and 73 perinuclear regions for 0 and 20 min, respectively, k = 3 embryos. (F) Normalized intensity of apical MTs at 0 and 20 min; n = 396 and 377 regions for 0 and 20 min, respectively, k = 3 embryos. (G) Stable perinuclear MT pools indicated by brightly stained acetylated MT (green) around nuclei (DAPI, magenta). (H) Still frames showing FRAP of α-tubulin:GFP at centrosomes (top) and perinuclear region (bottom). Time is indicated in seconds; photobleaching was performed at 0 s. Circle or rectangle in magenta indicates the photobleached region. (H′) Fluorescence recovery profile for centrosome and perinuclear regions. (H′′) Halftime of recovery for centrosomal vs perinuclear α-tubulin:GFP (mean ± SEM). (H‴) Immobile fraction for centrosomal vs perinuclear α-tubulin:GFP (mean ± SEM). For (H′–H‴), n = 11 FRAP regions for each plot, k = 11 embryos. Scale bar = 5 μm for (A′) and (G), 3 μm for (A) and (A′′), and 2 μm for (H). All scatter plots show the mean ± SD. Statistical significance was calculated using the Mann–Whitney U-test. **P < 0.01 and ****P < 0.0001.

MTs undergo a centrosomal to non-centrosomal transition during GBE. (A–C) MTs marked with Jupiter:GFP at level of centrosomes 20 min pre-GBE (A), at GBE onset (B), and 20 min into GBE (C). Arrows point to centrosomal MT. (A′–C′) Perinuclear MT pools (green) at 20 min pre-GBE (A′), at onset (B′), and 20 min into GBE (C′). (A′’–C′′) Orthogonal projection of MT and nuclei in epithelium 20 min before GBE (A′′), at onset (B′′), and 20 min into GBE (C’’). (A‴–C‴) Line scan intensity plot for MT as denoted in the lines (blue = 2 μm above nuclear midplane, red = at nuclear midplane) shown in A′′, B′′, and C′′, respectively. (D) Normalized intensity of centrosomal MTs at 0 and 20 min GBE; n = 150 centrosomes for each time point, k = 3 embryos. (E) Normalized intensity of perinuclear MTs at 0 and 20 min GBE; n = 80 and 73 perinuclear regions for 0 and 20 min, respectively, k = 3 embryos. (F) Normalized intensity of apical MTs at 0 and 20 min; n = 396 and 377 regions for 0 and 20 min, respectively, k = 3 embryos. (G) Stable perinuclear MT pools indicated by brightly stained acetylated MT (green) around nuclei (DAPI, magenta). (H) Still frames showing FRAP of α-tubulin:GFP at centrosomes (top) and perinuclear region (bottom). Time is indicated in seconds; photobleaching was performed at 0 s. Circle or rectangle in magenta indicates the photobleached region. (H′) Fluorescence recovery profile for centrosome and perinuclear regions. (H′′) Halftime of recovery for centrosomal vs perinuclear α-tubulin:GFP (mean ± SEM). (H‴) Immobile fraction for centrosomal vs perinuclear α-tubulin:GFP (mean ± SEM). For (H′–H‴), n = 11 FRAP regions for each plot, k = 11 embryos. Scale bar = 5 μm for (A′) and (G), 3 μm for (A) and (A′′), and 2 μm for (H). All scatter plots show the mean ± SD. Statistical significance was calculated using the Mann–Whitney U-test. **P < 0.01 and ****P < 0.0001.

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