Figure S2. Patronin function contributes... | Rockefeller University Press

Figure S2.

$Patronin function contributes to the stabilization of perinuclear MT networks. (A) Quantification showing depleted MT intensities in apical regions above nuclei in Patronin-compromised embryos as compared to the control at 20 min into GBE; n = 200 regions for control and n = 287 regions for Patronin from k = 3 embryos for each background. (B) Still frames showing FRAP of α-tubulin:GFP at centrosomes (top) and perinuclear region (bottom) in Patronin embryos. Time is indicated in seconds; photobleaching was performed at 0 s. A circle or a rectangle in magenta indicates the photobleached region. (C) Fluorescence recovery profile for centrosome and perinuclear regions in Patronin embryos; n = 11 centrosomal regions from k = 11 embryos and n = 9 perinuclear regions from k = 9 embryos. (D) Halftime of recovery and immobile fraction for centrosomal α-tubulin:GFP in control and Patronin embryos (mean ± SEM); n = 11 centrosomal regions from k = 11 embryos for each background. (E) Halftime of recovery and immobile fraction for perinuclear α-tubulin:GFP in control and Patronin embryos (mean ± SEM); n = 11 and 9 perinuclear regions from k = 11 and 9 for control and Patronin, respectively. (F) Intensity heatmap of acetylated MT from immunostained control and Patronin embryos, revealing fragmented and destabilized MT at perinuclear regions when Patronin function is compromised. Arrows mark perinuclear acetylated MT. (F′) Quantitation of acetylated MT pools; n = 70 regions from k = 7 embryos from each background. (G) Patronin (magenta) localization transitions during GBE (0 and 20 min). (G′) Cortical Patronin:GFP intensities in region 2 μm below the apical cell surface at the onset and 20 min into GBE; n = 60 cells from k = 3 embryos for each background. (H) Patronin intensities at centrosomes at 0 and 20 min GBE; n = 150 centrosomal regions for each time point from k = 3 embryos. (I) Box plot showing peak nuclear speeds in control and Patronin embryos; n = 546 and 291 in control and Patronin, respectively, from k = 3 embryos for each background. Scale bar = 2 μm for (B), and 5 μm for (F and G). All scatter plots show the mean ± SD. Statistical significance was calculated using the Mann–Whitney U-test. ***P < 0.001 and ****P < 0.0001.$

Patronin function contributes to the stabilization of perinuclear MT networks. (A) Quantification showing depleted MT intensities in apical regions above nuclei in Patronin-compromised embryos as compared to the control at 20 min into GBE; n = 200 regions for control and n = 287 regions for Patronin from k = 3 embryos for each background. (B) Still frames showing FRAP of α-tubulin:GFP at centrosomes (top) and perinuclear region (bottom) in Patronin embryos. Time is indicated in seconds; photobleaching was performed at 0 s. A circle or a rectangle in magenta indicates the photobleached region. (C) Fluorescence recovery profile for centrosome and perinuclear regions in Patronin embryos; n = 11 centrosomal regions from k = 11 embryos and n = 9 perinuclear regions from k = 9 embryos. (D) Halftime of recovery and immobile fraction for centrosomal α-tubulin:GFP in control and Patronin embryos (mean ± SEM); n = 11 centrosomal regions from k = 11 embryos for each background. (E) Halftime of recovery and immobile fraction for perinuclear α-tubulin:GFP in control and Patronin embryos (mean ± SEM); n = 11 and 9 perinuclear regions from k = 11 and 9 for control and Patronin, respectively. (F) Intensity heatmap of acetylated MT from immunostained control and Patronin embryos, revealing fragmented and destabilized MT at perinuclear regions when Patronin function is compromised. Arrows mark perinuclear acetylated MT. (F′) Quantitation of acetylated MT pools; n = 70 regions from k = 7 embryos from each background. (G) Patronin (magenta) localization transitions during GBE (0 and 20 min). (G′) Cortical Patronin:GFP intensities in region 2 μm below the apical cell surface at the onset and 20 min into GBE; n = 60 cells from k = 3 embryos for each background. (H) Patronin intensities at centrosomes at 0 and 20 min GBE; n = 150 centrosomal regions for each time point from k = 3 embryos. (I) Box plot showing peak nuclear speeds in control and Patronin embryos; n = 546 and 291 in control and Patronin, respectively, from k = 3 embryos for each background. Scale bar = 2 μm for (B), and 5 μm for (F and G). All scatter plots show the mean ± SD. Statistical significance was calculated using the Mann–Whitney U-test. ***P < 0.001 and ****P < 0.0001.

This Feature Is Available To Subscribers Only