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Figure 1. A-E: In situ hybridization with the XSnail2 RNA probe on stage 18N.F. embryos into which 1 ng of mutant RhoGTPase mRNA was microinjected, into one blastomere, at stage 6 N.F. The injected side (left), marked with an asterisk, is compared with the noninjected (right) side; lysine-fixable fluorescein was injected together with mRNA and visualized by blue staining (black arrows). B-E: Embryos were injected with N19RhoA mRNA (B), V14RhoA mRNA (C), N17Rac1 mRNA (D), V12 Rac1 mRNA (E). F: NC territory of embryos scanned with ImageJ 1.36b software. The graph shows the ratio of stained NC areas on the injected side to NC areas on the noninjected side. G-K: In situ hybridization with the XFoxD3 RNA probe on stage 18N.F. embryos into which 140 pg of pCMV DNA constructs encoding RhoGTPase mutants was microinjected into one left dorsal blastomere at stage 3N.F. The injected side (left), marked with an asterisk, is compared with the noninjected (right) side; beta -galactosidase mRNA was co-injected and visualized by Red-Gal staining. H-K: Embryos were injected with pCMV-N19RhoA (H), pCMV-V14RhoA (I), pCMV-N17Rac (J), pCMV-V12 Rac1 (K). L: NC territory of embryos scanned with ImageJ 1.36b software. The graph shows the ratio of stained NC areas in injected sides to NC areas in noninjected sides. M-Q: In situ hybridization with the XSnail1 RNA probe on stage 18N.F. embryos into which 140 pg of pCMV DNA constructs encoding RhoGTPase mutants was microinjected, into one blastomere, at stage 3N.F. The injected side (left), marked with an asterisk, is compared with the noninjected (right) side; beta -galactosidase mRNA was co-injected and visualized by Red-Gal staining. N-Q: Embryos were also co-injected with N19RhoA mRNA (N), V14RhoA mRNA (O), N17Rac1 mRNA (P), V12 Rac1 mRNA (Q). R: NC territory of embryos scanned with ImageJ 1.36b software. The graph shows the ratio of stained NC areas on the injected side to NC areas on the noninjected side. n, total number of scanned embryos; CMV, cytomegalovirus. |
