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microscopy\nKidneys were dissected, fixed in paraformaldehyde, treated with methanol, and washed with PBS containing Tween-20 (PBS-T) prior to treatment with lectins and antibodies. PBS-T was used for incubations of the tissues with fluoroscein-conjugated Dilichos biflorus aggutinin (DBA, Vector; Burlingame, USA), and PBS-T plus 2% goat serum was used for incubations with the antibodies. The primary antibodies were anti-WT1 (c-19; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-uvomorulin (e-cadherin, Cdh1, Sigma-Aldrich St. Louis, MO, USA), and anti-Cited1 (Neomarkers-Lab Vision Corporation, Fremont, CA, USA). The secondary antibodies were Alexa 555-conjugated anti-rabbit and Alexa 633-conjugated anti-rat antibodies (Molecular Probes, Eugene, OR, USA).\nThe tissues were imaged with a laser scanning microscope (Carl Zeiss, Thornwood, New York, USA) equipped with an argon (488 nm) and two HeNe lasers (543 nm and 633 nm). Optical sections approximately 2 μm thick were obtained every 5 μm to a depth of at least 65 μm. The sections began at the surface of the kidney and were on a plane tangential to it. Two Z-stack series were obtained, one from each of the two kidneys of each embryo. Ureteric bud tips identified by section tracing were counted within a defined area of the confocal image."}

    craft-ca-core-ex-dev

    {"project":"craft-ca-core-ex-dev","denotations":[{"id":"T19093","span":{"begin":709,"end":719},"obj":"MOP:0000779"},{"id":"T19092","span":{"begin":688,"end":694},"obj":"NCBITaxon:9986"},{"id":"T19091","span":{"begin":672,"end":682},"obj":"MOP:0000779"},{"id":"T19090","span":{"begin":662,"end":671},"obj":"CHEBI:52673"},{"id":"T19089","span":{"begin":646,"end":656},"obj":"GO:0042571"},{"id":"T19088","span":{"begin":570,"end":576},"obj":"PR_EXT:000005505"},{"id":"T19087","span":{"begin":520,"end":524},"obj":"PR_EXT:000001447"},{"id":"T19086","span":{"begin":508,"end":518},"obj":"PR_EXT:000001447"},{"id":"T19085","span":{"begin":496,"end":506},"obj":"PR_EXT:000001447"},{"id":"T19084","span":{"begin":432,"end":435},"obj":"PR_EXT:000017459"},{"id":"T19083","span":{"begin":411,"end":421},"obj":"GO:0042571"},{"id":"T19082","span":{"begin":387,"end":397},"obj":"GO:0042571"},{"id":"T19081","span":{"begin":347,"end":352},"obj":"UBERON:0001977"},{"id":"T19080","span":{"begin":342,"end":346},"obj":"NCBITaxon:9925"},{"id":"T19079","span":{"begin":264,"end":281},"obj":"NCBITaxon:271171"},{"id":"T19078","span":{"begin":253,"end":263},"obj":"MOP:0000779"},{"id":"T19077","span":{"begin":241,"end":252},"obj":"CHEBI:31624"},{"id":"T19076","span":{"begin":228,"end":235},"obj":"UBERON:0000479"},{"id":"T19075","span":{"begin":194,"end":195},"obj":"CHEBI:53424"},{"id":"T19074","span":{"begin":178,"end":188},"obj":"GO:0042571"},{"id":"T19073","span":{"begin":139,"end":140},"obj":"CHEBI:53424"},{"id":"T19072","span":{"begin":125,"end":133},"obj":"CHEBI:53424"},{"id":"T19071","span":{"begin":84,"end":92},"obj":"CHEBI:17790"},{"id":"T19070","span":{"begin":53,"end":69},"obj":"CHEBI_EXT:paraformaldehye"},{"id":"T19069","span":{"begin":20,"end":27},"obj":"UBERON:0002113"},{"id":"T19103","span":{"begin":1212,"end":1229},"obj":"UBERON:0006364"},{"id":"T19102","span":{"begin":1204,"end":1210},"obj":"UBERON:0000922"},{"id":"T19101","span":{"begin":1188,"end":1195},"obj":"UBERON:0002113"},{"id":"T19100","span":{"begin":1084,"end":1090},"obj":"UBERON:0002113"},{"id":"T19099","span":{"begin":890,"end":895},"obj":"CHEBI_EXT:argon"},{"id":"T19098","span":{"begin":781,"end":788},"obj":"UBERON:0000479"},{"id":"T19097","span":{"begin":751,"end":757},"obj":"CHEBI_SO_EXT:molecular_probe"},{"id":"T19096","span":{"begin":741,"end":750},"obj":"CHEBI_EXT:polyatomic_entity_or_group"},{"id":"T19095","span":{"begin":729,"end":739},"obj":"GO:0042571"},{"id":"T19094","span":{"begin":725,"end":728},"obj":"NCBITaxon:10114"}],"text":"Confocal microscopy\nKidneys were dissected, fixed in paraformaldehyde, treated with methanol, and washed with PBS containing Tween-20 (PBS-T) prior to treatment with lectins and antibodies. PBS-T was used for incubations of the tissues with fluoroscein-conjugated Dilichos biflorus aggutinin (DBA, Vector; Burlingame, USA), and PBS-T plus 2% goat serum was used for incubations with the antibodies. The primary antibodies were anti-WT1 (c-19; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-uvomorulin (e-cadherin, Cdh1, Sigma-Aldrich St. Louis, MO, USA), and anti-Cited1 (Neomarkers-Lab Vision Corporation, Fremont, CA, USA). The secondary antibodies were Alexa 555-conjugated anti-rabbit and Alexa 633-conjugated anti-rat antibodies (Molecular Probes, Eugene, OR, USA).\nThe tissues were imaged with a laser scanning microscope (Carl Zeiss, Thornwood, New York, USA) equipped with an argon (488 nm) and two HeNe lasers (543 nm and 633 nm). Optical sections approximately 2 μm thick were obtained every 5 μm to a depth of at least 65 μm. The sections began at the surface of the kidney and were on a plane tangential to it. Two Z-stack series were obtained, one from each of the two kidneys of each embryo. Ureteric bud tips identified by section tracing were counted within a defined area of the confocal image."}

    craft-ca-core-dev

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