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Supplementary Material. Fig. S2 Pronephric phenotype induced by injection of MoPax2.2. Whole mount in situ hybridization for tubule and glomus marker genes. Embryos were injected with MoPax2.2 at the 4-cell stage, in the equatorial region of both left blastomeres. They were cultured until the tailbud stages 3335 (A) or 25 (B) and analyzed by in situ hybridization with specific probes for the indicated genes. For each embryo, injected (aq) and uninjected (aq) sides are shown. Pax2 depletion does not lead to any change in the expression of pronephric marker genes at stage 25. At the late tailbud stage, gene expression in the proximal part of the tubule, including nephrostomes, is affected by MoPax2.2 while expression in the distal tubule is not modified except for the terminal differentiation marker clcnkb1 whose expression is completely inhibited. Expression of the glomus marker genes wt1 and nephrin is not significantly affected in MoPax2.2 injected embryos. Insets show higher magnification of the anterior pronephric territory. Numbers indicate how many embryos showed the phenotype among the total number of injected embryos. Image published in: Buisson I et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue pax2.L LOC108697493, pax-2, pax2-a, pax2-b, XPax-2, XPax2 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric kidney pronephric nephrostome pronephric duct glomus optic stalk otic vesicle spinal cord midbrain-hindbrain boundary post-anal gut rgn.L regucalcin, smp30, xsmp-30, xsmp30 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 glomus pronephric kidney cimap1a.L odf3 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric nephrostome epidermis mecom.L ecotropic viral integration site 1, evi-1, evi1, mecom-a, mecom-b, xEvi-1 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric kidney pronephric duct pronephric tubule proximal tubule distal tubule forebrain midbrain hindbrain cranial neural crest slc4a4.L hhNMC, hnbc1, nbc1, nbc2, nbce1, pNBC, slc4a5, XNBC1 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 otic vesicle pronephric kidney pronephric duct pronephric tubule proximal tubule forebrain olfactory placode slc12a1.L Na-K-2Cl cotransporter, NKCC-2, NKCC2, XNKCC2 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric kidney pronephric duct pronephric tubule proximal tubule atp1b1.S atp1b1a, Beta1-Na/K-ATPase, Na+-K+-ATPase, β1-NaK-ATPase X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 otic vesicle olfactory placode lens intersomitic region pronephric kidney pronephric duct pronephric tubule proximal tubule distal tubule post-anal gut clcnkb.L ClC-k, clc-kb, clckb, clcnka, clk-k2, xClC-k X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric duct pronephric tubule proximal tubule distal tubule cldn19.S claudin-19 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 pronephric duct pronephric nephrostome nphs1.S nephrin X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 glomus wt1.S wt-1, wt1-a, wt1-b, Xwt-1, xwt1 X. laevis Sometime during NF stage 33 and 34 to NF stage 35 and 36 glomus pronephric tubule proximal tubule pax2.L LOC108697493, pax-2, pax2-a, pax2-b, XPax-2, XPax2 X. laevis Throughout NF stage 25 optic vesicle spinal cord lateral plate mesoderm dorsal lateral plate mesoderm pronephric mesenchyme post-anal gut hnf1b.L hnf-1beta, hnf1-beta, hnf1beta, lfb3, tcf2, vhnf1 X. laevis Throughout NF stage 25 pronephric mesenchyme posterior neural tube lhx1.L lim-1, Lim1, lim1, Xlim-1, Xlim1 X. laevis Throughout NF stage 25 pronephric mesenchyme spinal cord hindbrain mecom.L ecotropic viral integration site 1, evi-1, evi1, mecom-a, mecom-b, xEvi-1 X. laevis Throughout NF stage 25 pronephric mesenchyme forebrain-midbrain boundary forebrain telencephalon neural crest cranial neural crest mandibular crest hey1.S bc8, chf2, herp2, hesr1, hrt-1, hrt1, oaf1, XHRT1 X. laevis Throughout NF stage 25 glomus pronephric mesenchyme somite head somite trunk somite presomitic mesoderm caudal presomitic mesoderm optic vesicle neural crest cranial neural crest brain forebrain midbrain hindbrain glomeral mesenchyme wt1.S wt-1, wt1-a, wt1-b, Xwt-1, xwt1 X. laevis Throughout NF stage 25 glomus pronephric mesenchyme glomeral mesenchyme Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + pax2 MO3 + NF33/34-35/36 (in situ hybridization) Sup.Fig. S2 A a-k Expression Phenotype decreased amount atp1b1.S expression in proximal tubule decreased amount cimap1a.L expression in pronephric nephrostome absent clcnkb.L expression in pronephric duct absent clcnkb.L expression in pronephric kidney decreased amount cldn19.L expression in proximal tubule decreased amount mecom.L expression in proximal tubule decreased amount pax2.L expression in proximal tubule decreased amount rgn.L expression in glomus decreased amount slc12a1.L expression in proximal tubule Larger Image Printer Friendly View

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