FIG. 3.

PFO permeabilization facilitates the release of chlamydial proteins from the cytosol of infected cells. Supernatants were collected from mock-infected HeLa cells (Not Inf) treated with SDS (SDS) or C. trachomatis-infected HeLa cells (Infected) treated with either HBSS alone (HBSS), PFO-diluted in HBSS (PFO), or SDS (SDS). (A) Autoradiogram of samples harvested after infected monolayers were incubated in the presence of [³⁵S]methionine-cysteine and eukaryotic protein synthesis inhibitors. Samples were loaded into wells as follows: HBSS treated (100 μl/well), PFO treated (100 μl/well), or SDS treated (10 μl/well) and resolved by SDS-polyacrylamide gel electrophoresis. The gel was dried and exposed to film. (B) Western blot of samples after being probed with mouse anti-GAPDH (GAPDH), mouse anti-MOMP (MOMP), or mouse anti-CPAF (CPAF). Equal volumes of supernatants from all treatment groups were loaded in each well.