Decreased <i>Pax2</i> expression in <i>p53^−/−</i> embryonic kidneys.

<p><b>A</b>) p53 expression overlaps that of Pax2 in the developing kidney. Immunofluorescence staining was done on E15.5 kidney sections to visualize Pax2 and p53 protein expression. p53 co- localizes with Pax2 in the cap mesenchyme (CM) and in the ureteric tip (UB). <b>B</b>) In situ hybridization. Decreased expression of <i>Pax2</i> mRNA at E12.5. <b>C</b>) QPCR. <i>Pax2</i> mRNA is significantly lower in germ-line <i>p53^−/−</i> kidneys. QPCR was done on RNA from individual E15.5 kidney pairs from wild-type and <i>p53^−/−</i> embryos (n = 4). <b>D</b>) Pax2 protein in E15.5 kidneys, detected by IF staining. Both wild-type and mutant kidney sections were processed for IF staining simultaneously and images captured at identical exposure setting. After normalizing intensity to that in wild-type, images were converted to a heat-map to demonstrate differential Pax2 staining between wild-type and mutant kidneys. High intensity staining corresponds to red/yellow areas and lower intensity corresponds to green/blue. Top panels at x4 and bottom panels at x20. <b>E</b>) Quantitation of staining intensity was done using Slidebook software and is shown graphically. UB, ureteric bud/tips; CM, cap mesenchyme; Nn, nascent nephrons including pretubular aggregates and renal vesicles.