Expression data from WT, Crebbp+/- and Ep300+/- bone marrow stroma

We found that the bone marrow microenvironment of Crebbp+/- mice was unable to properly maintain the immature stem - and progenitor pools. Instead, it stimulates myeloid differentiation that progresses into a myeloproliferative-like disease. Since CREBBP is a transcriptional co-activator, we used gene expression analysis to globally assess functional deficiencies in Crebbp+/- bone marrow stroma cells at a molecular level. Ep300 encodes a protein which is highly similar in structure and function to CREBBP; nevertheless, Ep300+/- mice suffer neither excessive myeloid differentiation nor loss of HSCs. Therefore, to identify expression changes specifically related to Crebbp heterozygosity, we focused on genes that showed significant differences in expression levels between Crebbp+/- and wild-type bone marrow stroma but no difference between Ep300+/- and wild-type. Bone marrow stroma was established from wild-type, Crebbp+/- and Ep300+/- mice that were 3-4 months old for RNA extraction and hybridization on Affymetrix microarrays. There are 4 biological replicates for each genotype used.我们发现，Crebbp+/-小鼠的骨髓微环境无法正常维持未成熟干细胞与祖细胞库的稳态，反而会促进髓系分化，并最终进展为类骨髓增殖性疾病。鉴于CREB结合蛋白（CREBBP）是一种转录共激活因子，我们通过基因表达分析在分子层面全面评估了Crebbp+/-骨髓基质细胞的功能缺陷。Ep300编码的蛋白在结构与功能上与CREBBP高度相似；然而，Ep300+/-小鼠既不会出现过度髓系分化，也不会发生造血干细胞（Hematopoietic Stem Cells, HSCs）的丢失。因此，为了筛选出与Crebbp杂合性特异性相关的基因表达变化，我们将研究焦点集中在那些在Crebbp+/-与野生型骨髓基质细胞中表达水平存在显著差异，而在Ep300+/-与野生型细胞间无表达差异的基因上。我们从3-4月龄的野生型、Crebbp+/-及Ep300+/-小鼠中分离并培养骨髓基质细胞，用于RNA提取及Affymetrix基因芯片杂交。每种基因型均设置4个生物学重复样本。