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Figure 7


Fig. 7. Analysis of phosphohistone H3 and Islet1 double-positive cells in mouse embryos at the 5- to 7-somite stage, in left and right horns of the sinus venosus of embryos at the 7-somite stage and in 6-somite-stage posterior SHF explants. (A) Quantitative analysis of the percentage of proliferating cells in right (R) and left (L) sides of the SHF with respect to the total number of Islet1 (Isl1)-positive cells in this mesoderm at the 5-, 6- and 7-somite stages (s). (A') Percentage of proliferating cells with respect to the total number of cells in the right and left sides of the sinus venosus (sv) at the 7-somite stage both for wild-type and for Pitx2c-/- embryos. The values are calculated from ten sections, with two embryos analysed for each stage. Red arrowheads, P≤0.03; black arrowheads, P≤0.025; asterisks, P≤0.01. (B-E) Examples of immunofluorescence analysis on a section of a wild-type (B,C) and a Pitx2c-/- (D,E) embryo at the 7-somite stage. (B,D) Merge of phosphohistone H3 (PHH3) and Hoechst staining on a wild-type (B) and on a Pitx2c-/- (D) embryo section (the arrows indicate PHH3-positive cells within the sinus venosus). NT, neural tube. (C,E) Merge of PHH3 and Islet1 immunofluorescence (the same sections as shown in B and D). The arrowheads indicate Islet1 and PHH3 double-positive cells. En, endoderm; SHF, second heart field. The white lines in B and D indicate the separation of the right (R) and left (L) sides of the sinus venosus. (F-I) Explants of the left (F,G) and right (H,I) pSHF from a Pitx2c-/- embryo after 72 hours of culture. (F,H) Hoechst staining; (G,I) PHH3 immunofluorescence. (J) Quantitative analysis of PHH3-positive cells as a percentage of the total number of cells in ten explants from left (L) and right (R) sides of Pitx2c+/- and Pitx2c-/- embryos. White, percentage in left Pitx2c+/- explants; black, percentage in right Pitx2c+/- explants; grey, percentage in left Pitx2c-/- explants; dark grey, percentage in right Pitx2c-/- explants. Arrowheads, P≤0.03; asterisks, P≤0.02.





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