Journal: Frontiers in Immunology
Article Title: Hypoxia Exacerbates Inflammatory Acute Lung Injury via the Toll-Like Receptor 4 Signaling Pathway
Figure Lengend Snippet: Differential gene expression profiles were identified in the total cells of bronchi alveolar lavage fluid (BALF) by high-throughput microarray analysis. Before the gene chip detection, we mixed three animal samples into one pooled sample and total of three pooled samples were detected in each group. The clustering display was generated by Chip software with two-way data clustering. Each column represents an individual gene, and each row corresponds to an individual array. Gene expression values were standardized and color-coded relative to the mean (green, values less than the mean; red, values greater than the mean), the gene expression profiles of the COMB group are significant different from the other three groups (A) . We used the Scatter Plot to show the differential patterns of gene expression in the four groups (B) . We used a Venn diagram to show the distribution patterns of gene changes in lipopolysaccharides (LPS) group, HPO group, and COMB group (C) . Microarray findings of gene expression pattern were validated by using real-time quantitative PCR (RT-qPCR) (D) .
Article Snippet: The RT-qPCR reactions were conducted using a Bio-Rad real-time PCR system with the following modified program: initial denaturation at 95°C for 2 min; followed by 40 cycles of denaturation at 95°C for 30 s, annealing at 60°C for 30 s, and amplification at 72°C for 20 s. Products were verified by melting curve analysis and 1.5% agarose gel electrophoresis.
Techniques: Expressing, High Throughput Screening Assay, Microarray, Chromatin Immunoprecipitation, Generated, Software, Real-time Polymerase Chain Reaction, Quantitative RT-PCR