Fig. 4

Histologic evaluation using maturation markers validates the graft medulla as perinatal stage. (a) In situ hybridization of maturation markers for the CD (Aqp4 and Aqp2) at E15.5, P0, and P7. Wnt7b is used as a constant control. Scale bars: 500 μm. (b) In situ hybridization of the CD maturation markers in the P0 kidney and the day 12 graft. Scale bars: 500 μm. 2nd and 3rd rows: magnified images of the 1st row. Scale bars: 100 μm. Two grafts from two independent transplantation experiments were analyzed. (c) In situ hybridization of maturation markers for the iMS (Hif1a and Ptgs2) at E15.5, P0, and P7. Scale bars: 500 μm. 2nd row: magnified images of the medulla in the 1st row. Scale bars: 100 μm. 3rd row: magnified images of the cortex in the 1st row. Ptgs2 is also expressed in the macula densa of the cortex. Scale bars: 100 μm. (d) In situ hybridization of the iMS maturation markers in the P0 kidney and the day 12 graft. Scale bars: 500 μm. 2nd row: magnified images of the medulla in the 1st row. Scale bars: 100 μm. 3rd row: magnified images of the cortex in the 1st row. Ptgs2 is also expressed in the macula densa of the cortex. Scale bars: 100 μm. Two grafts from two independent transplantation experiments were analyzed. (e) In situ hybridization of neuronal genes at E15.5, P0, and P7. Scale bars: 500 μm. 2nd row: magnified images of the medulla and cortex in the 1st row. Cntn6 is expressed in the oMS at E15.5. Scale bars: 100 μm; 3rd row: magnified images of the cortex in the 1st row. Gria1 is expressed in stromal progenitor at E15.5 and P0. Scale bars: 100 μm. (f) In situ hybridization of neuronal genes in the E15.5 kidney and the day 12 graft. Scale bars: 500 μm. 2nd row: magnified images of the medulla and cortex in the 1st row. In the graft, Cntn6 is expressed in the oMS, and Gria1 is expressed in the iMS and oMS. Scale bars: 100 μm; 3rd row: magnified images of the cortex in the 1st row. Scale bars: 100 μm. Two grafts from two independent transplantation experiments were analyzed.