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Fig. 4. Sox3, Sox2 and Sox3-VP16 inhibit Xvent2 expression and epidermal formation. (A) WISH for epi-k and Xp63 in embryos injected with 400–800 pg of sox3 or sox2 mRNA with lacZ (100 pg) mRNA into one of 2-cells. Uninjected (UI) side in top row, injected (I) side in bottom with arrow indicating injected area. Anterior is to the left and dorsal to the top. (B) Sox3, sox3-EnR, or sox3-VP16 mRNA (200–400 pg) was injected with lacZ mRNA into one of 2 blastomeres and gastrula or neurula embryos were assayed for bmp4, Xvent2 or epi-k expression by WISH. Bmp4 and Xvent2 (st. 12) embryos are animal pole view with the dorsal side to the top and epi-k embryos (st. 17) are a lateral view with anterior to the left. (C) WISH of stage 12 embryos injected with Sox3-GR into one blastomere of 2-cell embryos and untreated or treated with Dex at stage 9.5. Embryos are an animal pole views with dorsal to the top. (D) Diagram of BMP positive feedback loop, BRE-luciferase construct and relative luciferase activities in bar graph form. One-cell embryos were injected in the animal pole with vector or BRE-luciferase with sox3, sox2, sox11, sox3-EnR (400 pg), or sox3-VP16 mRNA (200 pg) and 50 pg CABR mRNA (top: lanes 3–9, and bottom: 2–5). The graph represents one experiment done in triplicate. Horizontal lines represent standard deviations. *p < 0.05 indicates values that differ significantly from lane 2 and **p < 0.01 indicates values that differ significantly from lane 6, Student’s t-test. |
