FIG 5.

The ATP level and protein-bound NAD(P)H fluorescence lifetime decreased after tanshinone IIA treatment. (A) Quantification of the relative amount of ATP. HEp-2 cells were infected with 10 IFUs of C. pneumoniae (Cpn) and treated with or without tanshinone IIA (TIIA; 25 μM). Relative ATP levels were measured directly (0 hpi) and at 6 hpi and 12 hpi. (B and C) HEp-2 cells were infected with C. pneumoniae, and tanshinone IIA was applied in parallel, followed by incubation for 48 h. (B) Gray-scale images of the NAD(P)H fluorescence intensity (top) and color-coded images of the protein-bound NAD(P)H fluorescence lifetime (τ₂-NAD(P)H) (bottom) in C. pneumoniae-infected cells not treated (left) and treated (right) with tanshinone IIA. Dashed lines and arrowheads indicate representative chlamydial inclusions which were selected for quantitative analysis. ns, nanoseconds. (C) Quantitative analysis of τ₂-NAD(P)H inside the chlamydial inclusion. Chlamydial inclusions (n = 30) from three different samples were analyzed on two independent experimental days. Data, presented as the mean ± SEM, are representative of those from three independent experiments (n = 3). *, P < 0.05.