RNA levels for the genes of interest were normalized to 36B4 expression level, whose CT values are represented in the upper row of the Table. All RNA values in the infected cells are referenced to non-infected control Mevastatin reverses LDL-receptor mRNA decline Inhibitors of HMG-CoA reductase are the most powerful activators of LDL receptor, whose activity on the LDL-receptor is mediated by SREBP pathway [21]. The addition of mevastatin to HepG2 cells infected with C. trachomatis at MOI of 1 did not affect cell viability nor

mRNA levels of 36B4 (Table 2). However, LDL-receptor mRNA level was dose-dependently upregulated with the increasing concentrations of mevastatin, reaching 2 fold induction at 40 μM level. This effect was even more

pronounced at 72 hours of the post-infection period though cell viability was declining (results not shown). NVP-AUY922 chemical structure There is also dose-dependent upregulation of cholesterologenic enzymes (HMG-CoA reductase, HMG-CoA synthase, SS) which is well known effect of statins in the cultures cells [22]. Selleck Napabucasin Notably, LDL-receptor related protein mRNA was not impacted under all conditions studied. Table 2 Folds and mRNA changes in C. trachomatis -infected HepG2 cells after addition of mevastatin.     Infected cells — Addition of mevastatin Parameter Non-infected cells 0 μM 1 μM 20 μM 40 μM 36B4ct 16.94 17.04 16.94 16.98 17.01 HMG-CoA Red 1 1.06 1.17 1.7 1.81 HMG-CoA Synth 1 0.79 1.46 1.53 1.89 SS 1 0.87 1.27 1.54 1.73 LDL-R 1 Suplatast tosilate 0.69 1.38 1.63 2.08 LRP 1 1.09 0.85 0.91 0.99 FAS 1 0.95 0.92 0.89

0.96 HepG2 cells were set up, grown and infected with C. trachomatis in presence or absence of mevastatin as described in Methods. RNA was extracted in 48 hours after inoculation of the bacteria. RNA levels for the genes of interest were normalized to 36B4 expression level, whose CT values are represented in the upper row of the Table. All RNA values in the infected cells are referenced to non-infected control. Mevastatin inhibits chlamydial growth in HepG2 cells Figure 1 shows representative immunofluorescent images of HepG2 cells infected with C. trachomatis in presence of increasing concentrations of mevastatin. As can be seen, the effect of mevastatin was marginal at the concentration of 1 μM, though some decline in the number of infected cells has been noticed. However, 20 μM mevastatin reduced both the number of inclusion bodies in the infected cells, promoting a perinuclear selleck chemicals pattern of staining. Mevastatin-treated cells (20 μM) appeared to contain smaller inclusion bodies similar to those that occur during persistent chlamydial infection [23]. The highest concentration of mevastatin tested (40 μM) abolished the number of infected cells almost completely. Analysis of bacterial transcripts showed a similar tendency.