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. 2017 Feb 15;7:40. doi: 10.3389/fcimb.2017.00040

Figure 1.

Figure 1

Induction of expression of APEX2 constructs from transformed Ctr L2. Ctr L2 was transformed with anhydrotetracycline (aTc)-inducible chlamydial expression vector pASK-L2 vector containing APEX2 (A), IncATM-APEX2 (TM = transmembrane domain only) (B), or IncF-APEX2 (C). HeLa cells were infected as described in Materials and Methods, and expression of constructs was induced 7 h post-infection with the indicated concentrations of aTc. Infected monolayers were fixed in methanol and processed for indirect immunofluorescence to detect expression of the construct with anti-FLAG antibody (red), the inclusion membrane with an anti-IncA antibody (green), or chlamydial organisms with an anti-Ctr L2 antibody (blue). Details for primary antibodies used in this study are found in Table 2. After incubation with the appropriate secondary antibodies, coverslips were mounted. Images were taken with Olympus Fluoview 1000 Laser Scanning Confocal Microscope (60x magnification with 2x zoom). Scale bars equal 10 μm; “a” indicates inclusions with aberrant bacteria; white arrows point to foci formed by secreted IncF-APEX2; the white asterisk indicates small inclusion formed by organisms expressing IncF-APEX2.