Figure 2. Effect of anhydrotetracycline on C. trachomatis.

L929 cells were infected with wild-type C. trachomatis L2 and then incubated in the presence of various concentrations of ATc throughout the infection (0 to 24 hpi). Cultures were fixed at 24 hpi and stained for MOMP (green). Representative images from three different concentrations are shown (A). Additionally, the same samples (and additional samples) were analyzed using Cell Profiler to calculate the average inclusion size (B). Over 700 inclusions were analyzed at each concentration. Separately, infected L929 cell cultures were treated with different concentrations of ATc and lysed at 24 hpi. Lysates were used to infect a fresh monolayer (in the absence of ATc) and infection levels were enumerated at 24 hpi using immunofluorescence microscopy. Percent inhibition of progeny was determined relative to a lysate from an untreated control (C).