Streptococcus gallolyticus subsp. gallolyticus promotes colorectal tumor development
HCT116, HT29, or FHC cells (~1x105/well) were incubated with L. lactis or TX20005 (~1x105/well) or media only for 12 hours. Cells were pulsed with 10 μM BrdU for 30 mins, incubated with anti-BrdU antibodies and secondary antibodies, and analyzed by flow cytometry (A—C). The level of PCNA was determined by western blot assays using total cell lysates from cells co-cultured with TX20005, L. lactis or media only (D—F). Representative images are shown. Band intensity was quantified using Image J, normalized to β-actin, and combined from at least three independent experiments (G–I). Apoptotic cells were detected by staining cells with PI and anti-Annexin V antibodies and secondary antibodies, followed by flow cytometry (J–L). Each experiment was done with duplicate wells and was repeated at least three times. Data are presented as mean ± SEM. One-way, two tailed ANOVA followed by SNK test was used for statistical analysis. *, p < 0.05; **, p < 0.01.