Journal:

Article Title: Reversal of the Antichlamydial Activity of Putative Type III Secretion Inhibitors by Iron ▿

doi: 10.1128/IAI.00023-07

Figure Lengend Snippet: Expression of T3S genes analyzed throughout the developmental cycle of C. trachomatis serovar D using RT-PCR, as well as expression of three chlamydial genes, groEL, ompA, and omcB, that have been shown to correlate with the early, middle, and late developmental cycle, respectively (26). The gene for the 16S ribosomal protein (CT026) was also included as an internal control for quantitation. As indicated at the top, HeLa 229 cells infected with C. trachomatis serovar D were grown in the presence of 20 μM INP 0406, INP 0341, or Desferal (400 μM) and harvested at 4 and 24 h postinfection (hpi). The RNA was isolated, and the equivalent of 5,000 genomic copies of C. trachomatis was used in an RT-PCR with primers for the genes indicated on the left (Table ​(Table1).1). For each time point both the RT-PCR results (+) and the results without added RT (−), to control for DNA contamination, are shown. The DMSO-treated samples yielded the same gene expression pattern as INP 0406 (data not shown). Amplicons were separated on a 2% agarose gel and stained with ethidium bromide. The TriDye 2-Log DNA ladder (0.1 to 10.0 kb) (New England Biolabs) was used as markers (lane M). Lane PCR contained a control for the PCR within the RT-PCRs. ORF, open reading frame.

Article Snippet: The TriDye 2-Log DNA ladder (0.1 to 10.0 kb) (New England Biolabs) was used as markers (lane M).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitation Assay, Infection, Isolation, Agarose Gel Electrophoresis, Staining