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. 2011 Nov 3;23(1):27–35. doi: 10.1681/ASN.2011010027

Figure 1.

Figure 1.

Nephrin immunostaining patterns in glomeruli of newborn CRE[+];NOTCH-IC mice after podocyte-specific removal of Rbpj. Shown are images of mouse glomeruli after triple indirect immunofluorescence labeling of newborn kidney tissue sections with anti-nephrin (green), anti-WT1 (red), and anti-MYC (blue) antibodies. (A–C) Serial images of a representative glomerulus from a newborn CRE[−];NOTCH-IC (control) mouse. (D–F) Serial images of a representative glomerulus from a newborn CRE[+];NOTCH-IC mouse. (D'–F') Magnified view of region denoted by white boxes in D–F, respectively. (G–I) Serial images of a representative glomerulus from a newborn CRE[+];NOTCH-IC;RBPKO mouse. (G'–I') Magnified views of region denoted by white boxes in G–I, respectively. (A, D, D', G, and G') Green and red channel images depicting immunostaining patterns for anti-nephrin and anti-WT1, respectively. (B, E, E', H, and H') Green and blue channel images depicting immunostaining patterns for anti-nephrin and anti-MYC, respectively. (C, F, F', I, and I') Merged green, red, and blue channel images. (A–C) Glomeruli of CRE[−];NOTCH-IC mice show anti-nephrin staining along the basolateral aspect of WT1-stained podocytes (A) and do not exhibit anti-MYC staining (B). (D–F) White boxes in D–F denote a glomerular segment in which anti-nephrin staining is absent. Light blue arrows in E and E' denote MYC-positive cells, which do not stain positively for nephrin. Of note, some MYCNOTCH-IC–expressing cells retain nephrin expression as indicated by white arrow in E. Pink arrows in F and F' show that anti-MYC–positive cells are also positive for WT1 and are thus MYCNOTCH-IC–expressing podocytes. Merged image shows co-immunodetection of anti-MYC and anti-WT1 in cells (indicated by purple arrows), which also lack anti-nephrin staining. White arrow in F denotes a [MYC,WT1]–double positive cell stained positively for nephrin. (G and H) Nephrin staining is present in WT1-stained podocytes of CRE[+];NOTCH-IC;RBPKO mice (G) with podocyte-specific deletion of Rbpj. Light blue arrows in H and H' demonstrate MYC-positive cells, which also invariably stain positively for nephrin. Pink arrows in I and I' show corresponding [MYC,WT1]–double positive cells stained positively for nephrin.