FIG 3.
Effect of AFR incubation time over the flow cytometric enumeration of transconjugant (A) and donor (B) cell counts originating from anaerobic filter matings. Transconjugant and donor counts were detected via flow cytometry for a total of 50,000 bacterial events. Detection was performed hourly for 3 h and a final measurement was taken after overnight (ON) incubation. Filter matings were carried out overnight inside an anaerobic chamber, using a 1:1 donor/recipient ratio. E. coli MG1655 lacIq::mCherry was used as the donor harboring the pKJK5-gfpmut3 plasmid, and E. coli MG1655 as the recipient strain. Matings were performed in triplicate, and error bars indicate the standard errors of the mean. AFR was performed in phosphate buffer (pH 8) in the presence of chloramphenicol (100 μg/ml). Means with different letters are significantly different (analysis of variance [ANOVA]-Tukey's multiple-comparison test; P values of <0.0005 [***] and <0.00005 [****]).