Fig. 2. Rho1 protein is ubiquitously expressed, but concentrated apically, in blastoderm embryos. (A,B) Confocal micrograph showing Rho1 localization at interphase (A) and metaphase (B) in an early syncytial blastoderm stage. Note that the accumulation of Rho protein (arrows) relative to nuclei (arrowheads) changes with cell cycle phase. (C-D') Confocal micrograph showing P1D9 (C,C') and DE-cadherin (D,D') staining of embryos at the cellular blastoderm stage. Note apical accumulation of Rho1 protein at this stage (arrow in C'). (E-F) Rho protein is localized cytoplasmically. (E) Double staining with P1D9 (green) and propidium iodide (red) to label nuclei. (F) Double staining with P1D9 (green) and an antibody against Lava lamp (red), a protein present in the cytoplasm. (G) P1D9 staining of Rho1^E3.10 homozygous embryos at the cellular blastoderm stage. Note the lack of apical Rho1 accumulation (arrow, compare with C'). (H) Close-up of an embryo at the cellular blastoderm stage showing basal accumulation of Rho1 protein in cells underlying the pole cells (arrow). (I-J') P1D9 staining in stage 11 (I,I') and stage 14 (J,J') embryos. (I',J') Higher magnification views of the portion of the embryos boxed in I and J, respectively. Note subcellular punctate spots of Rho1 accumulation (arrows in I'), and that Rho1 protein does not accumulate in cells at the leading edge during dorsal closure (J'). (K) Neural commissures of a stage 14 embryo stained with P1D9. (L) Grazing section of a cellular blastoderm stage embryo showing accumulation of Rho1 protein at a puncture wound site. In all images except (L), anterior is left and dorsal is up. Scale bars: A,C',I': 10 µm; C, I: 50 µm; H,J',K,L: 25 µm.

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