Gene expression profile of uterine fibroblasts cocultured with epithelial cells infected with Chlamydia trachomatis [Coculture_scseq]

To identify potential downstream effects of infection-associated expression of pro-fibrotic signal factors, we have cocultured infected epithelial cells with uninfected fibroblasts, using single-cell RNA-sequencing to identify changes in gene expression associated with infection-associated paracrine signaling. Immortalized endocervical epithelial cells (End1/E6E7) seeded at 35% of confluence in a 6-well plate were either mock-infected or infected with Chlamydia trachomatis serovar R2 at an MOI of 2. At 1 hpi, infected epithelial cells were washed with heparan sulfate to remove transiently adhered organisms. Immortalized uterine fibroblasts (KCO2s) were then added into the same well. Cells were harvested for library preparation at 24 hpi (23 hours of coculture).