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Figure 2. Cell division is not required for cardiomyocyte differentiation. Embryos stained for phH3 expression after incubation without (A) or with HUA (B) from stage-13. (C) Percentage of phH3-positive cells in the epidermis are decreased after HUA treatment vs. untreated control (matched areas, n ≥ 3 separate embryos ± SD; *P < 0.001). Transverse sections of stage-33/34 embryos after incubation of embryos in normal media (D and D′) or media containing HUA (E and E′), Tropomyosin expression (red), phH3 (green), and DNA in nuclei (Hoechst, blue); §Myotome is also positively labelled by tropomyosin. Note: Stage-29/30 embryos after control (F and H) or HUA (G and J) incubation showing TIc (F and G) or Nkx2.10 (H and J) expressions. (K) Heart size in control or HUA-incubated embryos as measured by TIc and Nkx2.10 expression area (see Figure 4A–H for examples). |
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Figure 5. The requirement for Xic1 in cardiomyocyte differentiation is distinct from its ability to arrest the cell cycle. Stage-29/30 embryos either uninjected (A and H) or injected with CTRMo (B and I), Xic1Mo (C, J, and P), or Xic1Mo plus FL-Xic1 (30 pg) (D, K, and Q), NT-Xic1 (15 pg) (E and L), CT-Xic1 (30 pg) (F and M), Xic1(35–96) (30 pg) (G and N), Xic1CK- (R), or CTRMo plus Xic1CK- (S) detecting either Nkx2.10 (A–G) or TIc (H–S). (T) Percentage of stage-29/30 embryos with heart sizes 1–3 (cf. Figure 4) following injection with Mo and mRNA combinations as described. n = 26 – 53 embryos/treatment group. |
