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Tbx5 is required for chamber formation [34] and was normally expressed in Tbx3 mutant embryos (data not shown).
Expression of otog was unaffected in eis te296f mutant embryos (data not shown).
A similar change was detected in He sall mutant embryos (data not shown).
However, we have not observed changes in mespa/b expression in aplnrb mutant embryos (data not shown).
Upon examination of integrin α5-null nodes, we found ectopical distribution of F-actin in one out of three mutant embryos (data not shown), suggesting that additional FN1-binding integrins regulate apico-basal polarity of the nodal cells.
Furthermore, we found that 24% (13/54) of the Cas9/gRNA-treated embryos had the cardia bifida phenotype at 60 hpf as in fau tm236a mutant embryos (data not shown), suggesting efficient biallelic conversion of gata5 in the resulting somatic cells.
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Rib development was also normal in double-mutant embryos (data not shown).
mRNA levels of Shh, Fgf8 and Nkx2.1 were each significantly reduced in the Cdo –/− ;Boc –/− embryonic heads (Fig. 6); in similar analyses, Ptch1 and Gli1 mRNA levels were not significantly different between control and double-mutant embryos (data not shown).
However, we observed synthetic lethality in Dcas1/Dcas1; Src42adult08/CyO adult flies (Table 1, Figure 2A, C) and Dcas1/Dcas1; Src42Ak10108/Src42Ak10108 embryos, as fewer than 46% of these double mutant embryos hatched (data not shown).
Expression of Msx2, another marker of AVC myocardium, was downregulated in Tbx3 mutant embryos at E9.5 (data not shown).
We did not observe any differences in morphology or volume of the outflow tract or atrioventricular cushions in Robo1, Robo2, Slit2, or Slit3 mutant embryos at E14.5 (data not shown).
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