Fig. 8.

Cx43 carboxy-terminal is recruited to the nucleus by BTF3. a St24 embryos showing neural crest migration by ISH of twist. Arrowheads show normal and brackets impaired migration. Asterisk indicates the eye. Scale bar = 40 μm. b Neural crest migration index for the indicated treatment. (n_CMO = 89, n_BTF3MO = 98, n_{BTF3MO-BTF3FL} = 105, N = 4. c Lateral view of embryos at st24 showing n-cad expression, analyzed by ISH. Arrows show normal and brackets impaired expression, asterisk indicates the eye. d % of n-cad expressing embryos shown in c (n_CMO = 92, n_BTF3MO = 127, n_btFL+BTF3MO = 98 embryos, N = 4). e Analysis of N-cadherin expression in blastulae embryos (st9). Scale bar = 30 μm. f n-cad expression levels, representative embryos in e (n_CMO = 56, n_Cx-20k = 112, n_{Cx-20k-BTF3MO} = 88 embryos, N = 4). g Temporal analysis of Cx43-20k-GR nuclear localization in st23 neural crest after addition of dxm (time is shown in minutes after dxm treatment). Cx43-20k-GFP is shown in color-coded intensity and n-RFP is used to visualize the nucleus. Graph shows Cx43-20kGR-GFP nuclear fluorescence normalized to the cytosolic fluorescence (n_CMO = 25, n_BTF3MO = 32 explants, N = 3). h St23 neural crest, showing color-coded intensity of Cx43-20k-GFP compared with nRFP to visualize nuclear localization. In red diagrams BTF3 NLS deletion constructs are shown. Chart of nucleus vs. cytosol ratio of Cx43-20k-GFP (n_CMO = 25, n_BTF3MO = 39, n_BTF3MO-btf3 = 58, n_{BTF3MO-btf3ΔNLS} = 58, N = 3 independent experiments). Scale bars in g and h = 20 μm. In b box plots show the median, box edges represent the 25th and 75th percentiles, and whiskers show spread of data including outliers (Mann Whitney test p^*** < 0.001). Histograms in d, f, h, and lines in g represent mean and error bars S.E. (one-way ANOVA; p < 0.001; two-tailed t test p^** < 0.01, p^*** < 0.001. N number of independent experiments; n sample size. Spread of data in bar charts is shown as overlying dots. n.s. nonsignificant