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Figure 2


Fig. 2. Ncc migration defect in Sox3 null mouse embryos. (A-F) CRABPI immunofluorescence and 3D reconstruction. (A,B) Sox3 null embryo at 10 ss: only confocal sections through the region containing migrating ncc were used for 3D reconstruction (to avoid foregut diverticulum background). (A) Right side with PA2 migrating ncc. (B) Left side with ncc accumulating in front of PA2 (compare distance covered by ncc between both sides, arrows). (C) Wild-type and (D) Sox3 null embryo at 9.5 dpc, with ncc accumulating in front of PA2 (arrow). Staining is present in the distal part of the arch. (E) Wild-type and (F) Sox3 null embryo at 10.5 dpc with hypoplastic PA2. The bulge of ncc observed at 9.5 dpc is strongly reduced. (G,H) Dlx2 in situ hybridisation on 10.5 dpc wild-type (G) and Sox3 null embryos (H): reduction of Dlx2 staining in proximal PA2 (arrow). (I,J) PAAs staining by ink injection in wild-type (I) and Sox3 null (J) 9.5 dpc embryos, where PAA2 is missing. (K) Apoptosis quantification. An average of 42.7±1.95 CRABP/TUNEL-positive cells were present on the left side of affected embryos, whereas we counted 22.45±2 on the right side (P<0.0001, n=7). (L-O) TUNEL assay and CRABPI immunofluorescence. Coronal section of 9.5 dpc Sox3 null embryo affected on the left side only. DAPI (L). TUNEL assay (M). CRABPI (N). TUNEL/CRABPI merge picture (O). More apoptotic ncc are present on the left side of the embryo. Scale bar: 75 µm in L for L-O. a1-3, pharyngeal arch arteries 1-3; as, aortic sac; da, dorsal aorta.





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