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Fig. 4. XHRT1 early expression in the developing glomus is affected by translational inhibition of xWT1. (A) Design of the xWT1 MOs that target both pseudoalleles. (Bottom) In vivo translation of 500 pg of xWT1-eGFP is specifically inhibited by 15 ng of xWT1 MOs. (B-I) Control and injected sides of xWT1-depleted embryos stained with the indicated probes. xWT1 MOs abolished the expression of nephrin but had no effect on XSMP-30, Pax8 and Evi1. (J,K) Transversal sections of Evi1- and Pax8-stained xWT1-depleted embryos. Expression of both markers is unaffected by xWT1 knockdown. (L-O) Control and injected sides of xWT1-depleted embryos stained with XHRT1. Note that xWT1 knockdown decreases early but not late XHRT1 pronephric expression. |
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Fig. 6. Comparison of the ability of hGR/Su(H)/Ank and XHRT1-MT-hGR to affect the expression of proximal tubule and glomus markers. (A) Activation of Notch signaling until stage 25 expands xWT1 efficiently, while later activation had no effect. XSMP-30 expression was increased in some embryos activated between stages 22 and 27. Embryos were co-injected with hGR/Su(H)/Ank andβ -galactosidase mRNA. Injected embryos were treated, or not, with Dex at the indicated times and analysed at stage 32-35. Changes in the expression of xWT1 and XSMP-30 at each stage were scored in individual embryos by comparing the injected and injected sides in at least two different injections. Embryos were classified into three phenotypes (no changes, increase or decrease). n, number of cases analysed. (B-M) Control and injected sides of embryos injected with the indicated mRNA together with β-galactosidase mRNA, treated with Dex at stage 18 (xWT1, nephrin) or 25 (XSMP-30) and analysed with the indicated probes. Note that hGR/Su(H)/Ank expands xWT1, nephrin and, in a few cases, XSMP-30, while XHRT1-MT-hGR has no effect. |
