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Figure 4. Depletion of lhx1 results in a loss of the pronephric kidney.Embryos were injected (1xV2) with 300 pg of ctrl-AS or lhx1-AS at the 8-cell stage. (A, D, G, J) Uninjected embryos. (B, E, H, K) Embryos injected with ctrl-AS. (C, F, I, L) Embryos injected with lhx1-AS. (A–C) In situ hybridization of embryos at stage 20 for the pronephric marker pax8. (C) Reduced or absent pax8 expression was observed in 47% and 53% of the embryos, respectively (n = 34). (D–F) 3G8 whole-mount immunostaining was carried out at stage 32. (E) Reduced 3G8 staining was observed in 5% of the embryos (n = 41). (F) Reduced 3G8 staining was observed in 76% of the embryos (n = 34). Insets show enlargement of 3G8 staining of the proximal tubule. (G–I) 4A6 whole-mount immunostaining was carried out at stage 37/38. (H) Reduced 4A6 staining was observed in 9% of the embryos (n = 23). (I) Reduced 4A6 staining was observed in 91% of the embryos (n = 34). (J–L) In situ hybridization of embryos at stage 39 for β1-NaK-ATPase. (L) Reduced β1-NaK-ATPase expression was observed in 72% of the embryos (n = 29). (M) Bar graph with the percentage embryos injected with the different doses of lhx1-AS that showed presence, reduction or absence of pax8 expression at stage 20. Lhx1-AS 100 pg: reduced pax8 expression was observed in 89%, absence in 4% and presence in 7% of the embryos (n = 28). Lhx1-AS 200 pg: reduced pax8 expression was observed in 93% and presence in 7% of the embryos (n = 27). Lhx1-AS 300 pg: reduced of pax8 expression was observed in 47% and absent in 53% of the embryos (n = 34). Lhx1-AS 400 pg: reduced pax8 expression was observed in 50% and absent in 50% of the embryos (n = 42). Lhx1-AS 300 pg+zflhx1 mRNA 25 pg: presence of pax8 expression was observed in 51% of the embryos (n = 41). Image published in: Cirio MC et al. (2011) Image downloaded from an Open Access article in PubMed Central. This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication. Gene Synonyms Species Stage(s) Tissue pax8.L pax-8, XPax-8, XPax8 X. laevis Throughout NF stage 20 otic region intermediate mesoderm Image source: Published Larger Image Printer Friendly View

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