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analysis of Pygo1/Pygo2 mutant kidneys\nHistological examination of Pygo2 null and Pygo1/Pygo2 double-null mutant kidneys did not reveal any abnormalities in nephrogenesis (data not shown). Confocal analysis was therefore performed to characterize renal development in E18.5 Pygo1/Pygo2 null kidneys more precisely (Figure 3). Wt1 (red) and Cited1 (red) antibodies both stain the capping metanephric mesenchyme around the ureteric bud tips [17]. WT1 antibody also stains renal vesicles and glomerular anlage. Antibodies to Cdh1, also known as E-cadherin (blue), were used to identify epithelial structures, including the branching ureteric bud and nascent nephrons of the developing kidney [18]. Dolichos biflorus (DBA) lectin [16] was used to selectively stain ureteric bud-derived structures [19].\nFigure 3 Confocal analysis of Pygo1/Pygo2 mutant E18.5 kidneys. (B, D) Pygo1-/-/Pygo2-/- mutant E18.5 kidneys and (A, C) kidneys of normal littermates were stained using antibodies to (C, D) Cited1 or (A, B) Wt1, both expressed in the condensing metanephric mesenchyme, and colored as red, Cdh1, a general marker of epithelia (blue), and DBA lectin staining the ureteric tree [16]. Confocal Z-sections were obtained every 5 μm for 75–80 μm. A reduced number of ureteric tips per area was observed in (B, D) the Pygo1/Pygo2 null kidneys compared with (A, C) control littermates. In addition, ureteric bud tips were often more dilated in Pygo1/2 null mutants compared with controls. The condensed mesenchyme surrounding the ureteric bud was also significantly expanded in the mutants. Original magnification × 200. Wt1 and Cited1 (red) staining revealed an increase of approximately 30% in the thickness of the capping mesenchyme surrounding the mutant ureteric buds (Figure 3, Table 2). Nevertheless, the mutant metanephric mesenchyme underwent relatively normal nephrogenesis. Cdh1-staining nephrons (blue) were identified connecting to the ureteric bud tips [16] and extending into the medulla of the kidney, as normal. Intermediate structures of nephrogenesis, including renal vesicles, comma-shaped bodies, and S-shaped bodies appeared normal.\nTable 2 Confocal analysis of Pygo2 wild-type, heterozygous, and null E18.5 kidneys on the Pygo1 null background.\nGenotype Ureteric tip Condensing mesenchyme\nUreteric tips/mm2 SD n Mesenchyme condensate size (μm) SD n\nPygo2 +/+ 196 8.36 3 16.2 1.09 3\nPygo2 +/- 191 11.6 7 15.6 1.18 5\nPygo2 -/- 147* 12.2 7 20.9 * 1.12 5\n*Significant change (P \u003c 0.01) between the three groups.\nSD, Standard deviation; n, number Confocal analysis also showed that the ureteric bud tips [16] of Pygo1/Pygo2 null kidneys were dilated and misshaped compared with those of littermates with at least one wild-type Pygo2 allele (Figure 3). In addition, the Pygo1/Pygo2 double-homozygous mutant kidneys also had a decrease of approximately 25% in the number of ureteric bud tips per area compared with littermates with at least one wild-type copy of Pygo2 (Table 2). No significant difference in ureteric bud-tip density was seen between Pygo2+/- and Pygo2+/+ embryos (P = 0.33)."}

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analysis of Pygo1/Pygo2 mutant kidneys\nHistological examination of Pygo2 null and Pygo1/Pygo2 double-null mutant kidneys did not reveal any abnormalities in nephrogenesis (data not shown). Confocal analysis was therefore performed to characterize renal development in E18.5 Pygo1/Pygo2 null kidneys more precisely (Figure 3). Wt1 (red) and Cited1 (red) antibodies both stain the capping metanephric mesenchyme around the ureteric bud tips [17]. WT1 antibody also stains renal vesicles and glomerular anlage. Antibodies to Cdh1, also known as E-cadherin (blue), were used to identify epithelial structures, including the branching ureteric bud and nascent nephrons of the developing kidney [18]. Dolichos biflorus (DBA) lectin [16] was used to selectively stain ureteric bud-derived structures [19].\nFigure 3 Confocal analysis of Pygo1/Pygo2 mutant E18.5 kidneys. (B, D) Pygo1-/-/Pygo2-/- mutant E18.5 kidneys and (A, C) kidneys of normal littermates were stained using antibodies to (C, D) Cited1 or (A, B) Wt1, both expressed in the condensing metanephric mesenchyme, and colored as red, Cdh1, a general marker of epithelia (blue), and DBA lectin staining the ureteric tree [16]. Confocal Z-sections were obtained every 5 μm for 75–80 μm. A reduced number of ureteric tips per area was observed in (B, D) the Pygo1/Pygo2 null kidneys compared with (A, C) control littermates. In addition, ureteric bud tips were often more dilated in Pygo1/2 null mutants compared with controls. The condensed mesenchyme surrounding the ureteric bud was also significantly expanded in the mutants. Original magnification × 200. Wt1 and Cited1 (red) staining revealed an increase of approximately 30% in the thickness of the capping mesenchyme surrounding the mutant ureteric buds (Figure 3, Table 2). Nevertheless, the mutant metanephric mesenchyme underwent relatively normal nephrogenesis. Cdh1-staining nephrons (blue) were identified connecting to the ureteric bud tips [16] and extending into the medulla of the kidney, as normal. Intermediate structures of nephrogenesis, including renal vesicles, comma-shaped bodies, and S-shaped bodies appeared normal.\nTable 2 Confocal analysis of Pygo2 wild-type, heterozygous, and null E18.5 kidneys on the Pygo1 null background.\nGenotype Ureteric tip Condensing mesenchyme\nUreteric tips/mm2 SD n Mesenchyme condensate size (μm) SD n\nPygo2 +/+ 196 8.36 3 16.2 1.09 3\nPygo2 +/- 191 11.6 7 15.6 1.18 5\nPygo2 -/- 147* 12.2 7 20.9 * 1.12 5\n*Significant change (P \u003c 0.01) between the three groups.\nSD, Standard deviation; n, number Confocal analysis also showed that the ureteric bud tips [16] of Pygo1/Pygo2 null kidneys were dilated and misshaped compared with those of littermates with at least one wild-type Pygo2 allele (Figure 3). In addition, the Pygo1/Pygo2 double-homozygous mutant kidneys also had a decrease of approximately 25% in the number of ureteric bud tips per area compared with littermates with at least one wild-type copy of Pygo2 (Table 2). No significant difference in ureteric bud-tip density was seen between Pygo2+/- and Pygo2+/+ embryos (P = 0.33)."}

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analysis of Pygo1/Pygo2 mutant kidneys\nHistological examination of Pygo2 null and Pygo1/Pygo2 double-null mutant kidneys did not reveal any abnormalities in nephrogenesis (data not shown). Confocal analysis was therefore performed to characterize renal development in E18.5 Pygo1/Pygo2 null kidneys more precisely (Figure 3). Wt1 (red) and Cited1 (red) antibodies both stain the capping metanephric mesenchyme around the ureteric bud tips [17]. WT1 antibody also stains renal vesicles and glomerular anlage. Antibodies to Cdh1, also known as E-cadherin (blue), were used to identify epithelial structures, including the branching ureteric bud and nascent nephrons of the developing kidney [18]. Dolichos biflorus (DBA) lectin [16] was used to selectively stain ureteric bud-derived structures [19].\nFigure 3 Confocal analysis of Pygo1/Pygo2 mutant E18.5 kidneys. (B, D) Pygo1-/-/Pygo2-/- mutant E18.5 kidneys and (A, C) kidneys of normal littermates were stained using antibodies to (C, D) Cited1 or (A, B) Wt1, both expressed in the condensing metanephric mesenchyme, and colored as red, Cdh1, a general marker of epithelia (blue), and DBA lectin staining the ureteric tree [16]. Confocal Z-sections were obtained every 5 μm for 75–80 μm. A reduced number of ureteric tips per area was observed in (B, D) the Pygo1/Pygo2 null kidneys compared with (A, C) control littermates. In addition, ureteric bud tips were often more dilated in Pygo1/2 null mutants compared with controls. The condensed mesenchyme surrounding the ureteric bud was also significantly expanded in the mutants. Original magnification × 200. Wt1 and Cited1 (red) staining revealed an increase of approximately 30% in the thickness of the capping mesenchyme surrounding the mutant ureteric buds (Figure 3, Table 2). Nevertheless, the mutant metanephric mesenchyme underwent relatively normal nephrogenesis. Cdh1-staining nephrons (blue) were identified connecting to the ureteric bud tips [16] and extending into the medulla of the kidney, as normal. Intermediate structures of nephrogenesis, including renal vesicles, comma-shaped bodies, and S-shaped bodies appeared normal.\nTable 2 Confocal analysis of Pygo2 wild-type, heterozygous, and null E18.5 kidneys on the Pygo1 null background.\nGenotype Ureteric tip Condensing mesenchyme\nUreteric tips/mm2 SD n Mesenchyme condensate size (μm) SD n\nPygo2 +/+ 196 8.36 3 16.2 1.09 3\nPygo2 +/- 191 11.6 7 15.6 1.18 5\nPygo2 -/- 147* 12.2 7 20.9 * 1.12 5\n*Significant change (P \u003c 0.01) between the three groups.\nSD, Standard deviation; n, number Confocal analysis also showed that the ureteric bud tips [16] of Pygo1/Pygo2 null kidneys were dilated and misshaped compared with those of littermates with at least one wild-type Pygo2 allele (Figure 3). In addition, the Pygo1/Pygo2 double-homozygous mutant kidneys also had a decrease of approximately 25% in the number of ureteric bud tips per area compared with littermates with at least one wild-type copy of Pygo2 (Table 2). No significant difference in ureteric bud-tip density was seen between Pygo2+/- and Pygo2+/+ embryos (P = 0.33)."}

{"project":"2_test","denotations":[{"id":"17425782-2164159-8469451","span":{"begin":449,"end":451},"obj":"2164159"},{"id":"17425782-3902535-8469452","span":{"begin":699,"end":701},"obj":"3902535"},{"id":"17425782-15618518-8469453","span":{"begin":736,"end":738},"obj":"15618518"},{"id":"17425782-9639355-8469454","span":{"begin":803,"end":805},"obj":"9639355"},{"id":"17425782-15618518-8469455","span":{"begin":1969,"end":1971},"obj":"15618518"},{"id":"17425782-15618518-8469456","span":{"begin":2652,"end":2654},"obj":"15618518"}],"text":"Confocal analysis of Pygo1/Pygo2 mutant kidneys\nHistological examination of Pygo2 null and Pygo1/Pygo2 double-null mutant kidneys did not reveal any abnormalities in nephrogenesis (data not shown). Confocal analysis was therefore performed to characterize renal development in E18.5 Pygo1/Pygo2 null kidneys more precisely (Figure 3). Wt1 (red) and Cited1 (red) antibodies both stain the capping metanephric mesenchyme around the ureteric bud tips [17]. WT1 antibody also stains renal vesicles and glomerular anlage. Antibodies to Cdh1, also known as E-cadherin (blue), were used to identify epithelial structures, including the branching ureteric bud and nascent nephrons of the developing kidney [18]. Dolichos biflorus (DBA) lectin [16] was used to selectively stain ureteric bud-derived structures [19].\nFigure 3 Confocal analysis of Pygo1/Pygo2 mutant E18.5 kidneys. (B, D) Pygo1-/-/Pygo2-/- mutant E18.5 kidneys and (A, C) kidneys of normal littermates were stained using antibodies to (C, D) Cited1 or (A, B) Wt1, both expressed in the condensing metanephric mesenchyme, and colored as red, Cdh1, a general marker of epithelia (blue), and DBA lectin staining the ureteric tree [16]. Confocal Z-sections were obtained every 5 μm for 75–80 μm. A reduced number of ureteric tips per area was observed in (B, D) the Pygo1/Pygo2 null kidneys compared with (A, C) control littermates. In addition, ureteric bud tips were often more dilated in Pygo1/2 null mutants compared with controls. The condensed mesenchyme surrounding the ureteric bud was also significantly expanded in the mutants. Original magnification × 200. Wt1 and Cited1 (red) staining revealed an increase of approximately 30% in the thickness of the capping mesenchyme surrounding the mutant ureteric buds (Figure 3, Table 2). Nevertheless, the mutant metanephric mesenchyme underwent relatively normal nephrogenesis. Cdh1-staining nephrons (blue) were identified connecting to the ureteric bud tips [16] and extending into the medulla of the kidney, as normal. Intermediate structures of nephrogenesis, including renal vesicles, comma-shaped bodies, and S-shaped bodies appeared normal.\nTable 2 Confocal analysis of Pygo2 wild-type, heterozygous, and null E18.5 kidneys on the Pygo1 null background.\nGenotype Ureteric tip Condensing mesenchyme\nUreteric tips/mm2 SD n Mesenchyme condensate size (μm) SD n\nPygo2 +/+ 196 8.36 3 16.2 1.09 3\nPygo2 +/- 191 11.6 7 15.6 1.18 5\nPygo2 -/- 147* 12.2 7 20.9 * 1.12 5\n*Significant change (P \u003c 0.01) between the three groups.\nSD, Standard deviation; n, number Confocal analysis also showed that the ureteric bud tips [16] of Pygo1/Pygo2 null kidneys were dilated and misshaped compared with those of littermates with at least one wild-type Pygo2 allele (Figure 3). In addition, the Pygo1/Pygo2 double-homozygous mutant kidneys also had a decrease of approximately 25% in the number of ureteric bud tips per area compared with littermates with at least one wild-type copy of Pygo2 (Table 2). No significant difference in ureteric bud-tip density was seen between Pygo2+/- and Pygo2+/+ embryos (P = 0.33)."}