Expression data from Lbx1-EGFP+ flow sorted cells from Pitx2 WT, HT, Null forelimb tissue

Determing the gene network regulated by Pitx2 in during forelimb muscle development of mouse embryos at E12.5. Lbx1 is coexpressed in Pitx2+ cells during forelimb development, thus Pitx2-LacZ and Lbx1-EGFP+ mice were cross bred to allow us to purify Lbx1-EGFP+|Pitx2 -wildtype, het, or null cells by flow sorting We used microarray analysis to determine the genes misregulated in Pitx2 null mice compared to Pitx2 wildtype and heterozyogtes. Lbx1-EGFP+|Pitx2(+/+, LacZ/+, or LacZ/LacZ) cells were flow sorted, RNA extracted, labelled cDNA was hybridized to Affymetrix mouse genome 430 2.0 arrays本研究旨在确定小鼠胚胎E12.5时期前肢肌肉发育过程中受Pitx2调控的基因调控网络。Lbx1在前肢发育过程中与Pitx2阳性细胞共表达，因此我们通过将Pitx2-LacZ小鼠与Lbx1-EGFP阳性小鼠杂交，得以通过流式分选纯化出Lbx1-EGFP阳性且Pitx2为野生型、杂合或纯合缺失的细胞。我们采用基因芯片分析技术，对比Pitx2野生型及杂合型小鼠，筛选出Pitx2纯合缺失小鼠中相较于野生型和杂合型小鼠表达失调的基因。我们对Lbx1-EGFP阳性且Pitx2基因型为(+/+、LacZ/+或LacZ/LacZ)的细胞进行流式分选，提取RNA并标记cDNA后，将其与Affymetrix小鼠基因组430 2.0芯片进行杂交。