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. 2008 Jun;19(6):1139–1157. doi: 10.1681/ASN.2007050596

Figure 7.

Figure 7.

Analysis of Pax2 expression in podocytes of CRE(+);NOTCH-IC transgenic mice. (A through H) Pax2 mRNA in situ hybridization in frozen sections of transgenic mouse kidneys. Shown are representative images from P7 (A, B, E, and F) and P28 (C, D, G, and H) mouse kidneys. (A through D) Low-power images of CRE(−);NOTCH-IC (A and C) and CRE(+);NOTCH-IC (B and D) mouse kidney cryosections. (E through H) Magnified views of CRE(−);NOTCH-IC (E and G) and CRE(+);NOTCH-IC (F and H) glomeruli enclosed by black boxes in corresponding top panels. At P7, Pax2 mRNA transcripts are not detected in glomeruli of CRE(−);NOTCH-IC (A and E) or CRE(+);NOTCH-IC (B and F) mouse kidneys. In contrast, at P28, Pax2 mRNA transcripts are detected in podocytes of CRE(+);NOTCH-IC glomeruli (H, black arrows). Also, Pax2 mRNA is detected in parietal glomerular epithelium (H, black arrowhead). (I through N) Dual immunofluorescence labeling of mouse kidney tissue sections with anti-MYC and anti-PAX2 antibodies. Shown are representative serial micrographs of glomeruli obtained from P28 CRE(−);NOTCH-IC (I through K) and CRE(+);NOTCH-IC (L through N) mouse kidneys imaged by multichannel fluorescence microscopy. (I and L) Green channel images showing anti-MYC immunodetection with Alexa488-conjugated secondary antibody. Green arrows, cells stained positively with anti-MYC antibody. (J and M) Red channel images showing anti-PAX2 immunodetection with Alexa594-conjugated secondary antibody. Red arrows and arrowheads, cells stained positively with anti-PAX2. Sections were counterstained with DAPI, imaged by blue channel, and merged onto corresponding green and red channel images. (K and N) Merged images of corresponding green, red, and blue channel images. (N, yellow arrows) Cells showing overlapping staining with anti-MYC and anti-PAX2 antibodies, identifying these cells as coexpressing MYCNOTCH-IC and PAX2. (K and N, red arrowheads) PAX2-positive cells that do not stain positively with anti-MYC antibody. Magnifications: ×200 in A through D; ×1000 in E through N.