31. For the comparison of CF18acvs CF4ab, 76 out of the 1446 differentially expressed unique genes are immune related genes, which contained 27 and 49 more highly expressed genes in CF18ac and CF4ab, respectively. Of the more highly expressed genes in CF18ac, the highest fold change was observed for AK235118 which belongs to the sys temic lupus erythematosus pathway, while interleukin 8 was the www.selleckchem.com/products/MDV3100.html most highly expressed genes in CF4ab with a fold change of 4. 93. Validation of the microarray results by real time quantitative RT PCR To validate the microarray results by quantitative Inhibitors,Modulators,Libraries RT PCR, we designed primers for four up regulated, four down regulated and two unchanged genes from the three comparisons of CF4abvs control, CF4acvs control and CF18acvs control.
In addition, MUC4 was also vali dated using the primers reported by Sargeant et al. Two commonly used reference genes, i. e. GAPDH and ACTB, were used in the validation. The primers Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries were designed to span introns to avoid the influence of DNA contamination. As shown in Table 2, the expres sion profiles of these genes detected by qRT PCR were consistent with those by microarray, which confirmed the reliability of our microarray data. Discussion In the present study, genome wide gene expression pro files of porcine IPEC J2 cells infected by three ETEC strains separately was stud ied using Agilent Porcine Oligo Microarray. Differences of gene expression profiles between cells with and without infection as well as among cells infected with different ETEC strains separately were presented.
To our knowledge, this is the first report about the remarked differential responses of porcine IEC cells to the infections of the three ETEC strains. After infection with F4ab, F4ac and F18ac ETEC Inhibitors,Modulators,Libraries sep arately, 2443, 3493 and 867 differentially expressed genes were identified in the IPEC J2 cells, respectively. Gene Ontology analysis of these three groups of genes revealed that they shared six biological process terms, of which five are involved in the cell cycle progression. This indicated that the infections of the three ETEC strains all affected cell cycle progression through bacter ial toxins or cyclomodulins. The genes induced by F4ab ETEC and F4ac ETEC shared the most bio logical process terms and pathways, which was consist ent with the similarity of the antigenic structures of F4ab and F4ac fimbrial antigen.
Both of them Inhibitors,Modulators,Libraries have the a epitopes formed by the conserved region of the major F4 fimbrial subunit FaeG. However, they also have their own specific GO terms. The specific GO terms of the F4ab ETEC induced genes are associated with catabolic processes, whereas those of the F4ac ETEC induced genes are associated with im mune Pazopanib solubility response, inflammatory response and response to wounding, and apoptosis. These results implied why F4ac is the most common antigenic variant of F4 fim briae causing piglet diarrhoea. Differentially expressed genes induced by ETECs are involved in some important pathways.