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Fig. S1

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ZDB-IMAGE-130830-12
Source
Figures for Dray et al., 2013
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Figure Caption

Fig. S1

Analyses of Convergence Extension, Apoptosis, and Cell Proliferation in itgα5mo;αVmo Embryos, Related to Figure 1

(A) Wild-type, fn1mo;fn1bmo, itgα5mo;αVmo and Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos were fixed at the tailbud stage and in situ hybridization for her1 was performed. We then measured the medial-lateral width of the tailbud at the level of the posterior tip of the notochord. The fn1mo;fn1bmo embryos (n=30) exhibited a 17.5% increase in width of the tailbud (unpaired t-test; p-value <0.05) which is consistent with previous observations [1]. In contrast, neither itgα5mo;αVmo (n=37) nor Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos (n=24) displayed a change in tailbud width relative to wild type (n=55) (p-value >0.05). Error bars show standard error. The fn1 and fn1b morpholinos have been previously described [2].
(B) Cell death was assayed in wild-type, itgα5mo;αVmo and Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos. We fixed embryos at the 13-somite stage and assayed apoptosis via TUNEL. Briefly, embryos were fixed overnight in 4% PFA in PBS at 4°C and stored in 100% methanol at 20°C. Embryos were rehydrated and permeabilized in 100% acetone at 20°C for 20 min followed by several washes in 0.5% Triton X-100 and 0.1% sodium citrate in PBS and then treated for TUNEL assay using the in situ Cell Death Detection Kit, POD (Roche, Basel, Switzerland) according to the manufacture’s instructions. There was no clear difference in apoptosis in itgα5mo;αVmo embryos (n=12) nor in Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos (n=10) compared to wild type (n=12).
(C) Cell proliferation was assayed in wild-type, itgα5mo;αVmo and Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos. We fixed embryos at the 13-somites stage and assayed for cell proliferation via immunohistochemistry for Phosphorylated Histone H3 (PHH3) as previously described [3]. Compared to wild-type (n=5), itgα5mo;αVmo embryos (n=5) exhibited a 36% decrease in cell proliferation (unpaired t-test, p-value <0.05). Proliferation in Tg(tbx6l:itgα5-RFP);itgα5mo;αVmo embryos (n=5) was no different than in itgα5mo;αVmo embryos (p>0.05). Scale bars equal 200 microns.

Acknowledgments
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