Figure 8.
Interaction of vinculin with adhesion complexes is required for its functions in INM and cell cycle. (A) Drawing showing an idealization of the positions occupied by the nucleus and the centrosomes along INM and cell cycle. (B) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with geminin·AG (green), Arl13b·RFP (cilia marker, magenta), and Cep152·GFP (centriole marker, green) and stained with phalloidin (grays) and DAPI (blue). The area encircled in the top panel is magnified in the lower panels, in which different combinations of channels are displayed for clarity. Scale bar = 2 μm. (C) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with geminin·AG (green), VCL·mCherry (magenta), and Cep152·GFP (green) and stained with N-cadherin (cyan). Four different geminin·AG+ cells with the nucleus at different apical-basal positions are shown. The areas encircled in the left panels are magnified in the right panels, in which different combinations of channels are displayed for clarity. Arrows indicate the centriole position in the VCL·mCherry channel. Scale bar = 10 μm. (D) Apical region images of NTs electroporated at stage HH18 for 24 h with H2B-RFP (magenta), Cep152-GFP (green or gray), and shControl or shVCL. Arrowheads point to internalized centrosomes. Scale bar = 10 μm. (E) Bar graph showing the percentage of internalized centrosomes (>2 μm from the apical membrane) in NTs transfection with shControl or shVCL. Mean ± SEM. t test, n = 15 sections for control and 10 for shVCL from three experiments using at least three embryos per condition. (F) Dot plot showing the distance from the apical membrane to the centrosome of all internalized centrosomes (>2 μm from the apical membrane) from the data shown in Fig. 8 F. The black bars indicate the median. t test. * = P < 0.05, ** = P < 0.01.

Interaction of vinculin with adhesion complexes is required for its functions in INM and cell cycle. (A) Drawing showing an idealization of the positions occupied by the nucleus and the centrosomes along INM and cell cycle. (B) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with geminin·AG (green), Arl13b·RFP (cilia marker, magenta), and Cep152·GFP (centriole marker, green) and stained with phalloidin (grays) and DAPI (blue). The area encircled in the top panel is magnified in the lower panels, in which different combinations of channels are displayed for clarity. Scale bar = 2 μm. (C) Images of transverse sections of chicken NTs transfected at stage HH12 for 24 h with geminin·AG (green), VCL·mCherry (magenta), and Cep152·GFP (green) and stained with N-cadherin (cyan). Four different geminin·AG+ cells with the nucleus at different apical-basal positions are shown. The areas encircled in the left panels are magnified in the right panels, in which different combinations of channels are displayed for clarity. Arrows indicate the centriole position in the VCL·mCherry channel. Scale bar = 10 μm. (D) Apical region images of NTs electroporated at stage HH18 for 24 h with H2B-RFP (magenta), Cep152-GFP (green or gray), and shControl or shVCL. Arrowheads point to internalized centrosomes. Scale bar = 10 μm. (E) Bar graph showing the percentage of internalized centrosomes (>2 μm from the apical membrane) in NTs transfection with shControl or shVCL. Mean ± SEM. t test, n = 15 sections for control and 10 for shVCL from three experiments using at least three embryos per condition. (F) Dot plot showing the distance from the apical membrane to the centrosome of all internalized centrosomes (>2 μm from the apical membrane) from the data shown in Fig. 8 F. The black bars indicate the median. t test. * = P < 0.05, ** = P < 0.01.

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