Fig. S9

Evaluation of the roles of tnc and tnw in the mitigation of cardia bifida, and of the efficiency and specificity of tnw MOs. (A) Knockdown of tnc with tnc-MO2 in s1pr2^as10 mutants increased the percentages of 26-ss embryos raised at 22.5°C with the Class II and Class III cardia bifida phenotype. (B) Knockdown of tnw with tnw-MO2 in s1pr2^as10 mutant embryos raised at 28.5°C partially rescued cardia bifida phenotypes at 24 hpf. Class I (a single heart tube) to Class II (cardiomyocytes either in close proximity or in contact) and Class III (separated cardiomyocytes). Statistical significance was determined using Student’s t-test. * indicates p <0.05, ** indicates p <0.01. (C) Diagram indicating the relative binding positions of two tnw-MOs in the 5′untranslated region of tnw mRNA. (D) Green fluorescence can be detected in embryos co-injected with CMV-tnwUTR-EGFP and tnw-5mm at 24 hpf. Green fluorescence was not observed in the majority of embryos co-injected with CMV-tnwUTR-EGFP and tnw-MO1, while green fluorescence was detected in 30% of embryos co-injected with CMV-tnwUTR-EGFP and tnw-MO2. (E) Percentage of embryos expressing EGFP following co-injection of CMV-tnwUTR-EGFP with tnw-5mmMO, tnw-MO1 or tnw-MO2. The error bars indicate the standard error.