Chromatine immunoprecipation of atonal binding sites in Drosophila L3 Brains

Atonal is a proneural transcription factor expressed in the Drosophila neuroblast cluster known as the inner proliferation center (IPC). To characterize the putative targes of Atonal in the IPC we have used an endogenously GFP-tagged version of atonal to immunoprecipate brain samples with anti-GFP antibodies L3 larvae of the Ato::GFP genotype where dissected to collect developing brains. The fixed sample was homogenized by sonication. A 10% of the sonicated volume was processed as input controls while the rest was Immunoprecipitated with antiGFP antibodies, purified and sequence.

Atonal是一种成神经转录因子（proneural transcription factor），在果蝇（Drosophila）被称为内增殖中心（inner proliferation center, IPC）的成神经细胞簇中表达。为了表征Atonal在IPC中的推定靶标，我们利用内源性GFP标记的atonal变体，通过抗GFP抗体对Ato::GFP基因型三龄幼虫的脑组织样本开展免疫沉淀实验：首先解剖该基因型的三龄幼虫以收集发育中的脑组织，将固定后的样品经超声进行匀浆处理。取10%的超声处理体积作为输入对照，剩余样品则用抗GFP抗体进行免疫沉淀，随后纯化并完成测序。