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Fig. S4. Normal segmentation of the Inversin-deficient pronephros. (A) Schematic diagram of segment-specific expression of in situ markers. PT, proximal
tubule; IT, intermediate tubule; DT, distal tubule. (B–K) After unilateral injection of Invs-Mo, whole-mount in situ hybridization was performed at stage 38. The
sodium/glucose cotransporter (SGLT-1K) is expressed in the proximal tubule and nephrostomes (B and C). The sodium bicarbonate cotransporter NBC1 is
expressed in the proximal tubule and early distal tubule (DT1) but not the intermediate tubule (D and E). Note that the gap between proximal and distal
expression was clearly visible in Invs-Mo–injected embryos. The sodium/potassium/chloride transporter NKCC2 is expressed in the intermediate segment (F and
G), the thiazide-sensitive NaCl cotransporter NCC is expressed in the distal tubule (H and I), and ClC-K is expressed in the intermediate segment (J and K).
Intermediate tubules appear smaller and less extended on the injected side (arrowheads in G and K). Brackets in J and K mark the boundaries of ventral
extension of the intermediate tubule. (L and M) The glomerular marker Nephrin is equally expressed on both sides. (N) Dorso-ventral extension of the pronephric tubule measured in embryos stained for ClC-K in millimeters is shown. Coinjection of Inversin mRNA rescued the defects of Invs-Mo (*P ≤ 0.001, rank
sum test; the error bars represent SEM) |
