Late suppression of WNT signaling rescues CPs and cardiomyocyte numbers in tmem88a morphants. (A-D) WISH for nkx2.5 (box) and ntl in representative 8-somite stage control (Ctrl) and tmem88a (88a) morphants, without or with (+dkk1) WNT inhibition. Retraction of the notochord occurs in heat-shocked embryos, but independently of MO injection. Heat-shock treatment was administered to 75% epiboly embryos for 1 hour. n>20 embryos analyzed per panel. (E) nkx2.5+ area normalized to control values. n=6 embryos measured per condition. (F-I) Cardiomyocyte nuclei in representative 48-hpf embryos from the tg(myl7:DsRed2-nuc) line. n>25 embryos analyzed per panel. (J) Quantification of total cardiomyocyte (CM) numbers. n=6 control hearts analyzed; n=9 hearts analyzed for all other conditions. (K-N) Brightfield images showing craniocardiac tissues in 48-hpf embryos. n>50 embryos analyzed per condition. **P<0.01; ns, not significant; error bars indicate s.d.
|