Figure S3: Related to Figure 3. GRP defects in pkd2 morphants (A, B) SEM analysis of co-MO (A) and Pkd2-MO (B) injected specimens reveal loss of ciliation and altered GRP cell morphology in morphants. Examples are representative for 6 specimens analyzed each. Scale bars represent 20 micrometers. (C-H) Unilateral absence of dnah9 (D), nodal1 (F) and dand5 (H) in left-injected embryos in comparison to internal control sides at stage 17. Control embryos (C, E, G) show wildtype expression levels. (I) SEM analysis of ciliation and cell morphology in dorsal explant of stage 17 specimen unilaterally injected with Pkd2-MO targeted exclusively to the lateral, sensory part of the GRP. Shown is a ventral view of the GRP broken transversally and revealing the deep tissue arrangements in the top half of the picture. Right (I’) and left (I’’) magnifications show both the sensory areas of the GRP. Note that the lateral GRP cells were absent on the MO-injected side, such that central GRP cells, which are characterized by posteriorly polarized cilia, directly bordered non-ciliated endodermal cells. Sensory GRP cilia highlighted with red arrowheads, central GRP cilia with green arrowheads. (J-L) Expression of nodal1 in lateral GRP cells of flow stage (st. 17) specimens following Tp (K, L) or 1% DMSO (J) treatment during mid to late gastrula stage (st. 11.5). nodal1 was lost (K’) or strongly reduced (L) after Tp treatment, without causing gastrulation defects. (M-P) Expression of nodal1 in lateral GRP cells of flow stage (st. 16-19) specimens following 25-200µM BAPTA-AM (N-P) or 0.05-1% DMSO (M, P) treatment during mid and late gastrula stages (st. 11.5- 13/14). nodal1 was lost (O) or reduced (N) after BAPTA-AM treatment. *p<0.05, **p<0.01, **p<0.001 Black arrowheads indicate reduction or lack of expression. a, anterior; e, endoderm; l, left; n, notochord; p, posterior; r, right; s, somites
Image published in: Vick P et al. (2018)
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