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Figure 4—figure supplement 2. Localization of Pard3 mRNAs using RNAscope.

(A-C) Representative images (Imaris 3D-rendering) of RNAscope in situ hybridizations for Pard3aa, Pard3ba, and Pard3ab in 48–50hpf Tg(Kdrl:eGFP) control embryos. Images show the whole pattern of expression of Pard3aa, Pard3ba, and Pard3ab mRNAs in the trunk region (z-projections). Anatomical regions are outlined with white dashed lines, and roughly annotated based on location and with the support of transmitted light images (not shown). The aorta is outlined with magenta dashed lines. DLAV: dorsal longitudinal anastomotic vessels, SC: spinal cord, NT: notochord, SAS: sub-aortic space, PCV: posterior cardinal vein, PD: pronephric duct. Scale bars = 10 µm. (D) Representative images (Imaris 3D-rendering) of RNAscope in situ hybridizations for Pard3aa, Pard3ba, and Pard3ab from trunks narrowed down to aortic and sub-aortic regions of 48–50hpf Tg(Kdrl:eGFP) embryos. The aorta is outlined with white dashed lines. The RNAscope signals were segmented into spots and classified based on their localization: in aortic endothelial and hemogenic cells (magenta spots) or in extra-aortic tissues (grey spots). Scale bars = 10 µm.

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