FIG 4.

Tanshinone IIA treatment decreases the chlamydial load and limits inclusion development. HEp-2 cells were infected with C. pneumoniae (1, 10, or 25 IFUs) and in parallel treated with (w/) or without (w/o) tanshinone IIA (TIIA; 25 μM). After 48 h the cells were harvested, followed by mRNA isolation. (A) Quantification of relative chlamydial load using qRT-PCR. AU, arbitrary units. (B) Infected cells (10 IFUs) with and without tanshinone IIA treatment were fixed with methanol after 48 h, immunostained by use of an Imagen chlamydia kit (green, chlamydial LPS; red, cytoplasm), and counterstained with DAPI (blue). (Insets) Magnified images of the boxed areas. (C) Aberrant inclusions were quantified in C. pneumoniae-infected cells with and without tanshinone IIA treatment. Data, presented as the mean ± SD, and immunofluorescence images are representative of those from at least three independent experiments (n = 3 to 4). ***, P < 0.001.