Expression data of NIH3T3 in G0 and G1 phases

NIH3T3 in the middle of G0 to G1 transion consists of the cells which is still staying G0 phase and the cells which enters G1. Monitoring the expressions of p27 and Cdt1 enables to distinguish these two; p27+/Cdt1+ cells as the cells in G0 phase and p27-Cdt1+ cells as G1 phase We sorted p27+Cdt1+ (G0) NIH3T3 cells and p27-Cdt1+ (G1) NIH3T3 cells in the middle of G0-G1 transition, 5 hours after serum addition following the serum addition using mVenus-p27K- and mCherry-hCdt1(30/120) . We compared the expression profiles of these NIH3T3 cells in G0 and G1 phases and identified the genes upregulated in G0 or G1 phases. The p27+Cdt1+ (G0) NIH3T3 cells and p27-Cdt1+ (G1) NIH3T3 cells were sorted by FACS for RNA extraction and hybridization on Affymetrix microarrays.

处于G0到G1转换中期的NIH3T3细胞，包含仍停滞于G0期的细胞与已进入G1期的细胞。通过监测p27与Cdt1的表达水平，可对这两类细胞进行有效区分：其中p27+/Cdt1+细胞为G0期细胞，p27-Cdt1+细胞为G1期细胞。我们使用mVenus-p27K-与mCherry-hCdt1(30/120)完成标记后添加血清，并于血清添加后的第5小时，对处于G0-G1转换中期的p27+Cdt1+（G0期）NIH3T3细胞与p27-Cdt1+（G1期）NIH3T3细胞进行分选。随后我们比较了这两类处于G0、G1期的NIH3T3细胞的基因表达谱，鉴定得到在G0期或G1期上调的差异基因。前述p27+Cdt1+（G0期）与p27-Cdt1+（G1期）NIH3T3细胞通过荧光激活细胞分选术（FACS）分取后，用于RNA提取，并在Affymetrix基因芯片上完成杂交检测。