Mild KD of fn1b, itgα4, or vcam1, but not fn1a rescues increased SA sprouting in tardbp−/−; tardbpl−/− mutants. (A) Mild KD of fn1a, fn1b, itgα4, or vcam1 does not affect SA growth in wild type embryos compared to ctr MO injected siblings. Images showing rescue results with normal SA sprout number in tardbp−/−; tardbpl−/− mutants injected with 0.33 mM fn1b, 0.5 mM itgα4, or 0.75 mM vcam1 MO. Note the morphological difference compared to ctr MO injected tardbp−/−; tardbpl−/− mutants and the similarity to 0.75 mM ctr MO injected wild type sibglings. fn1a KD tardbp−/−; tardbpl−/− mutants (0.75 mM MO) are not rescued. (B) Statistical analysis demonstrating that mild KD of fn1b, itgα4, or vcam1 can statistically significantly rescue increased sprouting of SA from the DA at 24 hpf in tardbp−/−; tardbpl−/− mutants compared to ctr MO injected tardbp−/−; tardbpl−/− mutants. One of three (fn1a, fn1b, itgα4 KD) or two (vcam1 KD) independent experiments with two clutches per experiment are shown, normality test and unpaired t-test, n ≥ 23. (C) Mutant tardbp−/−; tardbpl−/− and control embryos carry the Tg (fli1:EGFP)y1 transgene, SA of the trunk dorsal of the yolk sack extension are shown. (D) Representative image of 1 mM itgα4 MO and itgα5/itgαv MO injected sibling and tardbp−/−; tardbpl−/− mutant embryos (right panel). Note the normal number of SA of the itgα4 MO injected tardbp−/−; tardbpl−/− mutant. Arrows point to SA defects in wild type siblings injected with 0.5 mM itgα5 and 1 mM itgαv MO indicating lack of rescue activity. Scale bar 50 μm, anterior to the left.
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