(A) Schematic representation of the experimental design to study the separate and combined effect of Chlamydia and HPV on the centrosome. Control cells are RPE-1 cells transfected with an empty plasmid. “HPV cells” are RPE-1 cells that stably express HPV16 oncoproteins E6 and E7 (gray cell). “Chlamydia cells” are RPE-1 cells infected with Chlamydia trachomatis (yellow circle), while “HPV+Chlamydia” cells are RPE-1 cells that stably express HPV16 oncoproteins and that are infected with C. trachomatis. Individual centrosomes are presented as green dots.
(B) Control RPE-1 cells or HPV cells, grown on coverslips, were either mock-infected or infected with C. trachomatis L2 at an MOI of 3. Samples were fixed at 36 hours post infection (hpi). Centrosomes were visualized with antibodies to γ-tubulin (green) and centrin (red), host and chlamydial DNA was detected with DAPI (blue). Chlamydial inclusions are outlined with white dashed lines. Scale bar: 5 μm.
(C) The percentage of host cells with supernumerary centrosomes (n>2 centrosomes) from the four different conditions is shown. 100 cells were analyzed for each condition. For the samples with a Chlamydia infection, only infected cells were examined and quantified. Data are represented as mean ± SD (n=3); **P≤0.01 and ***P<0.001.