FIG 5.

The overexpression of CT813-FLAG from C. trachomatis L2 results in reduced transcription of early and midcycle genes. HeLa cells were infected with C. trachomatis L2 CT813-FLAG, CT226-FLAG, or CT483-FLAG transformed strains, or wild-type C. trachomatis L2, and either not induced or induced at 7 hpi (5 nM or 20 nM aTc). RNA and DNA collected from separate wells of a 6-well plate at 7, 16, 24, and 36 hpi were processed as described in Materials and Methods. Normalized RNA was reverse transcribed to cDNA, and inc expression was measured by quantitative PCR. cDNA (ng) was normalized to genomic DNA (ng) for three biological replicates (except for clpP2, n = 2), and then these data were plotted using GraphPad Prism 8.4.0. Transcript profiles of early gene euo (A), midcycle gene clpP2 (B), late gene omcB (C), an early expressed inc gene, incE (D), and a midcycle inc gene, incA (E), are shown. Normalized transcripts from wild-type and C. trachomatis L2 CT813-FLAG were log transformed and then analyzed for statistical significance using a paired two-tailed Student's t test. Samples that were statistically analyzed were wild type and CT813 at 16 hpi (for omcB only), 24 hpi (all genes), and 36 hpi (all genes). Asterisks denote statistical significance: **, P ≤ 0.01; *, P ≤ 0.05.