In order to identify genes involved in the development of inner ear hair cells, we investigated the role of the transcription factor Islet-class LIM-homeodomain (LIM-HD) 1 (Isl1) in the development of the mouse prosensory region. Isl1 was deleted using the Pax2-Cre system, and deletion of both alleles was verified using cochlea sections. Changes in the number of prosensory region cells were measured to determine the effect of Isl1 on the development of the mouse prosensory region. In order to test whether Isl1 formed a protein complex with Ldb1 and Gata3, co-immunoprecipitation experiments were performed in HEK293 cells using the Flag-tagged LIM-domain of Isl1, HA-tagged LID of Ldb1 and Myc-tagged C-terminal domain of Gata3. The expression of Gata3, Sox2, Jag1 and P27 proteins in the prosensory region were not affected in Isl1-/- prosensory cells. Thus, Isl1 did not form a protein complex with Gata3 through Ldb1 in the Isl1-/- cells. Our results suggest that Isl1 may be dispensable for the development of the mouse prosensory region.

中文翻译：

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转录因子Isl1对于小鼠感觉区域的发育是必不可少的。

为了鉴定参与内耳毛细胞发育的基因，我们研究了转录因子Islet类LIM-同源域（LIM-HD）1（Isl1）在小鼠感觉区域发育中的作用。使用Pax2-Cre系统删除Isl1，并使用耳蜗切片验证两个等位基因的删除。测量前感觉区域细胞数目的变化以确定Isl1对小鼠前感觉区域发育的影响。为了测试Isl1是否与Ldb1和Gata3形成蛋白质复合物，使用Isl1的Flag标记的LIM结构域，Ldb1的HA标记的LID和Myc的C标记的C末端结构域在HEK293细胞中进行了免疫共沉淀实验。 Gata3。在Isl1-/-感觉细胞中，Gata3，Sox2，Jag1和P27蛋白在感觉区域中的表达不受影响。因此，Isl1在Isl1-/-细胞中未通过Ldb1与Gata3形成蛋白质复合物。我们的结果表明Isl1可能是小鼠的感觉区域发展的必要条件。