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Figure 1. Over-expression of lhx1 induces expansion of the pronephric kidney. Embryos were injected (1xV2) with 200 pg of LL-VP16 mRNA at the 8-cell stage. (A) In situ hybridization of embryos at stage 20 for the early pronephric marker pax8. (A) Uninjected embryo. (B) Control and injected sides of the same embryo are shown. (C) Expansion of pax8 expression was observed in 83% of the embryos (n = 33). Kidney fields are highlighted with black brackets. (D) 3G8 and 4A6 whole-mount immunostaining were carried out at stages 32 and 37/38, respectively. (D, G) Uninjected embryos. (E, F, H, I) Control and injected sides of the same embryo. (F, I) Larger tubule epithelium with 3G8 was observed in 81% of the embryos (n = 32) and expanded intermediate and distal tubules with 4A6 were observed in 80% of the embryos (n = 30). (D) Insets show enlargements of 3G8 staining of the proximal tubule. (J) 3G8, 4A6 double whole-mount immunostaining of stage 42 embryos. (K) Magnification of the pronephric kidney of uninjected embryo. (M) Magnification of the pronephric kidney of injected embryo. (N) Bar graph showing the percentage of embryos injected at the different doses of LL-VP16 that showed expansion of pax8 expression. LL-VP16 mRNA 50 pg: expansion of pax8 expression was observed in 35% of the embryos (n = 26). LL-VP16 mRNA 100 pg: expansion of pax8 expression was observed in 53% of the embryos (n = 30). LL-VP16 mRNA 200 pg: expansion of pax8 expression was observed in 70% of the embryos (n = 30). LL-VP16 mRNA 300 pg: expansion of pax8 expression was observed in 93% of the embryos (n = 27). |