VEGFA Upregulates FLJ10540 Figure 1. FLJ10540 exhibits differentially expressed patterns in lung adenocarcinomas, metastatic tumors, and a series of human lung invasive cancer cell lines. (CL1-0, CL1-1, CL1-5, and CL1-5-F4, in the order of increasing invasive activity). (A, left panel) The microarray expression patterns of FLJ10540 in lung adenocarcinoma patients were normalized against the expression patterns on HG_U133A chips. N: adjacent non-tumor tissues; T: tumor tissues. The boxplot shows the data distribution as a grouping classification and indicates that there is a statistically significant difference (p,0.0001) between the tumor tissues and the adjacent nont-umor tissues from the same lung cancer patient. (A, right panel) The mRNA levels of FLJ10540 in samples from 16 lung cancer patients were determined by Q-RT-PCR. The results were normalized against the expression levels of GAPDH mRNA in each paired sample and plotted with boxplot. (B) The microarray expression patterns of FLJ10540 were compared among 30 benign tumors, 20 tumor metastases to the lymph node, and 200 metastatic tumors, and were showed with boxplot. (C, left panel) The mRNA levels of FLJ10540 were determined by Q-RT-PCR in lung cancer cell lines. The results were normalized against the level of GAPDH mRNA in each cell line. The experiments were performed in triplicate. (C, right panel) Total protein was extracted from CL1-0 and CL1-5-F4 cells and subjected to immunoblot analysis with anti-FLJ10540. b-actin was used an internal loading control. doi:10.1371/journal.pone.0005052.g001 in a panel of cell lines, CL1-0, CL1-1, CL1-5, and CL1-5-F4, with either low or high invasive abilities, as previously described [23] by Q-RT-PCR. The data indicated that levels of FLJ10540 mRNA were high in CL1-5 and CL1-5-F4 cells that have high invasive and metastatic abilities, but were lower in the low invasive CL1-0 cells (Figure 1C, left panel). FLJ10540 protein levels were also markedly PLoS ONE | www.plosone.org 3 April 2009 | Volume 4 | Issue 4 | e5052