Figure 6. The block in C. trachomatis (Ct) inclusion growth through the inhibition of ACSLs is not dependent on lipid droplets (LD).

TC (A), 2-FPA and RG (B) were added to control and T863 treated DGAT2−/− MEF cells at two different concentrations (TC: 5 μM and 7.5 μM; 2-FPA: 200 μM and 300 μM; RG: 50 μM and 100 μM) for 16 h. The cells were then infected with Ct L₂ without removing the inhibitor from the media. After 24 hpi, samples were prepared for WB analysis. Membranes were probed with anti-Ct HSP60 antibody and anti-human GAPDH antibody as a loading control. (C) DGAT2−/− MEF cells were treated with the inhibitor T863 (lower panels) or left untreated (upper panels). After 8 h of incubation, the following ACSL activity inhibitors were added: 100 μM RG, 300 μM 2-FPA, and 7.5 μM TC. After 16 h of incubation, cells were infected with Ct L₂, without removing the inhibitors from the media. After 24 hpi, cells were fixed and visualized by confocal microscopy. Cells were stained with anti-Ct MOMP antibody (red), BODIPY 493/503 for neutral lipids (green), and Hoechst for nuclear and bacterial DNA (blue). Scale bar, 10 μm.