Figure 8

DsNLSm, defective in the β-catenin pathway, is active in noncanonical signaling. (a) Fz8-dependent recruitment of Dsh-GFP constructs to the cell membrane. Dsh-GFP or DsNLSm RNA (0.5 ng) was injected alone or with Fz8 RNA (1 ng) into two animal blastomeres at the 4–8-cell stage. GFP fluorescence was assessed in animal cap explants as in Figure 1b-d. Both Dsh and DsNLSm are efficiently recruited to the cell membrane by Fz8. Arrowheads point to cell membranes. (b) DsNLSm can rescue convergent extension defects caused by Xdd1. Four-cell embryos were injected with 0.6 ng Xdd1 RNA alone or together with 2 ng Dsh-GFP or DsNLSm RNA into two vegetal dorsal blastomeres. The injected embryos were allowed to develop until the sibling embryos reached stage 32. (c) Activation of JNK by the Dsh nuclear import mutant. Four animal blastomeres of four-cell embryos were each injected with 1 ng of RNAs encoding Dsh-GFP or DsNLSm. Embryonic lysates were collected at stage 10.5 for in vitro JNK activity assay using anti-phospho-specific c-Jun antibodies. Total GST-c-Jun levels were assessed with anti-GST antibodies. Dsh-GFP and DsNLSm were equally expressed, as monitored with anti-Dvl2 antibodies; β-tubulin served as a loading control.