Xenbase Image ID: 76229

FIG. 3. Induction experiments in Xlim-1-mutant-injected animal caps. Two-cell stage embryos were injected with 250 pg/embryo Xlim-1 mutant (Xlim-1/3m, Xlim-1-enR). (A) Immunohistochemistry of activin/retinoic acid-treated animal caps. All animal caps were cotreated with activin and retinoic acid for 3 h. Xlim-1-enR inhibited pronephros differentiation in a dose-dependent manner. Animal caps were bleached in H2O2 for 6 h after staining. (A) Differentiation of the pronephros was detected by the pronephric tubule-specific 3G8 antibody in control activin A/retinoic acid-treated animal caps. Arrows indicate signals (dark blue) of 3G8. In animal caps injected with 50 pg of Xlim-1-enR (B) and animal caps injected with 250 pg of Xlim-1-enR (C) the frequency of pronephros differentiation (41% in this experiment) was markedly decreased compared to animal caps injected with 50 pg of Xlim-1-enR (72%). (D) Differentiation patterns in Xlim-1 construct-injected animal caps. (D) Xlim-1/3m-injected untreated animal cap. Cement gland was observed (arrow). (E) Xlim-1/3m- injected animal cap with activin treatment. Arrows indicate notochord (Nc) and neural (N) tissues in animal cap. (F) Animal cap (injected with Xlim-1-enR) treated with activin A and retinoic acid. Though muscle and mesenchyme tissues were recognized (arrows), no pronephros was detected. (G) Pronephros (arrows) was recognized in Xlim-1/3m and Xlim-1-enR co-injected animal caps treated with activin A and retinoic acid. Cg, cement gland. N, neural tissue. Nc, notochord. M, muscle. Mes, mesenchyme. Pt, pronephric tubule. (H) Overexpression of Xlim-1 induced no pronephros marker genes. Lanes 1: Xlim-1/3m-injected explants. There is no expression of pronephric marker genes detected in Xlim-1/3m-injected explants with activin A (lane 1) or retinoic acid (lane 2) or in controls (lane 3). Lane 4: activin A/retinoic acid cotreated explants. Lane 5: sample from the whole embryo. Image published in: Chan TC et al. (2000) Copyright © 2000. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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