HCoV-229E infection in A549 cells: gene regulation in cells adjacent and distant to infected cells

In A549 cells, infection with human Coronavirus 229E spreads lateraly through the cell layer. This offers the opportunity to compare the gene expression patterns of infected cells with those of nearby unifected cells and to analyze paracrine effects occuring during infection. The aim of the experiment is to identify and compare changes in host gene expression of infected cells, cells immediately adjacent to infected cells and more distant cells with uninfected cells. To separate the different cell population, a Leica LMD6000 system was used. Individual cells were stained for the expression of the viral N protein or for DNA polymerase II by a shortened immunfluorescence protocol. After drying, the cell populations were visualized by fluorescence microscopy and separated by microdissection, collected in tubes and RNA was isolated on a small scale by column purification. Single-color hybridization of RNA from laser-microdissected cells. Duplicates.