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Nfection for experiments examining development, as presented in Fig. 7. Microscopy. HeLa monolayers were grown on 12-mm-diameter glass coverslips for immunofluorescence analyses. Monolayers were directly infected with Chlamydia followed by transfection with appropriate plasmid DNAs. Unless otherwise indicated, samples were routinely fixed with methanol at ca. 24 h postinfection for immunofluoresce
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Centrifugation, the pellets were washed three times, centrifuged, and resuspended in 200 l of buffer containing a 1 mM concentration of the substrate. After 60 min, the samples were centrifuged, and the absorbance of the supernatants was measured at 405 nm in an enzyme-linked immunosorbent assay (ELISA) reader. Control values (bacteria incubated in buffer) were used as a blank. No endogenous prote