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. 2009 Apr 13;106(18):7485–7490. doi: 10.1073/pnas.0811944106

Fig. 1.

Fig. 1.

Misexpression of Shh, Nodal, and Pitx2 on the right side does not affect sidedness of Tbx18 expression. (A, E, and I) Schematic depiction of the site of implantation of (A) Shh-expressing cells, (E) beads soaked in Nodal protein, or (I) Pluronic gel containing antisense Cfc oligonucleotides. Embryos were cultured until HH stage 12/13 and subjected to whole mount in situ hybridization analysis of (B-D, J, K, and P) Pitx2 (red color) and Tbx18 (blue color) expression. Red and blue arrowheads point to Pitx2 and Tbx18 expression domains, respectively. Only the heart region is shown in a ventral view, anterior end up. Embryos were treated on the right side of Hensen's node at HH stage 4 with (B-D) Shh-expressing cells, or (J and K) Cfc antisense oligonucleotides. (F and G) Whole mount in situ hybridization analysis of Nodal expression in HH stage 8 embryos after (F) control treatment or (G) implantation of Nodal protein at HH stage 6/7. (H) Tbx18 expression in a Nodal-treated embryo cultured until HH stage 12. (L) Proportion (%) of experimental embryos displaying aberrant Tbx18 or Nodal expression domains subsequent to Shh, Nodal, or antisense CFC treatment. Red bars indicate the percentage of embryos with aberrant Pitx2 expression, and yellow bars indicate the amount of normal left-sided expression. In the case of Nodal implantation, Nodal expression within LPM was analyzed. Blue bars display percentage of embryos with right-sided Tbx18 expression. (M-P) Electroporation of eGFP and Pitx2-RCAS constructs. (M-O) GFP fluorescent signal of an electroporated embryo at (M) HH stage 4, (N) HH stage 7, and (O) HH stage 12. (P) Forced expression of Pitx2 did not affect Tbx18 expression in the right sinus horn.