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. 2021 Jan 15;9(1):173. doi: 10.3390/microorganisms9010173

Figure 4.

Figure 4

Interfering with cytoplasmic [K+] leads to Chlamydia persistence. HeLa cells were infected with C. trachomatis LGV2. (A) Cells were treated with glibenclamide 12 hpi prior to fixation at 24 hpi followed by labelling with anti-Chlamydia (green) and DNA probe (DRAQ-5, magenta). Insert shows a normal inclusion (*) and abnormal inclusion (+) at higher magnification. Scale bar: 25 µm. (B) upper panel: schematic of the experiment design. I: infection. Lower panel: after treatment with glibenclamide and sample collection as described in the upper panel, lysate were tested for infectivity by measuring inclusion forming units per mL (IFU/mL). (C) Upper panel: schematic of the experiment design. I: infection. Lower panel: reversion of persistence assay, performed as described in the upper panel. Infectivity assays were performed (IFU/mL: inclusion forming unit per mL). For (B) and (C), error bars: standard deviation. (D) At 12 hpi, cells were treated with 1 µM nigericin, valinomycin or doxycycline. At 50 hpi, cells were prepared for RNA expression analysis by RT-PCR using 18S as a loading control and 16S as a marker of bacterial viability.