Estradiol clearly induced an overall down-regulation of chlamydial fatty acid biosynthesis, with seven genes being down-regulated at least 2-fold (accB, fabF, lipA, fabG, lplA_2). Estradiol also resulted in a marked down-regulation of the genes involved in chlamydial nucleotide (purine and pyrimidine) metabolism (adk, dnaE, dut, nrdA, surE, yggV, rpoC, ygfA, dut). In addition, we also observed a more minor down-regulation in cofactor and vitamin metabolism AZD5153 research buy pathways (hemC, hemN-1, yggV and folD). Table 3 Categorisation of the up- and down-regulated genes into pathways, as per KEGG.   Total Up-regulated

Down-regulated     Estradiol Progesterone Estradiol Progesterone Energy metabolism 14 3 4 6 4 Carbohydrate metabolism 23 2 9 1 – Lipid metabolism 27 1 2 7 8 Nucleotide Rabusertib molecular weight metabolism 29 – 1 16 3 Amino acid metabolism 30 3 8 3 3 Metabolism of other amino acids 4 – - – - Metabolism of cofactors and vitamins 33 – 1 6 3 Glycan biosynthesis and metabolism 16 2 6 1 2 Biosynthesis of secondary metabolism 15 1 1 3 4     12 32 43 27 The numbers represent the number of pathways (not

genes) affected following exposure with either Estradiol or progesterone. Taken together, this overall down-regulation of key pathways is suggestive of a persistence phenotype. The normal chlamydial developmental cycle can be altered under stressful conditions, leading to the formation of aberrant bodies (ABs) which are inhibited in their differentiation back to infectious EBs [11]. Molecular consequences include a ‘blockage’ in development involving down-regulation of late gene products in persistent infections [19]. The omcB and trpB genes are currently the most reliable general markers of chlamydial persistence [12–14, 20–22]. The down-regulation trends reported Orotidine 5′-phosphate decarboxylase in this project, for these genes under

estradiol supplement, were consistent with previous data in the microarray study of IFN-γ-mediated C. trachomatis serovar D persistence [13]. It has EPZ015938 previously been shown that trpA and trpB are two genes known to be involved in chlamydial persistence [12, 20]. Hogan et al. [12] showed that the expression patterns of these two genes were mostly up-regulated in chlamydial persistence. While the expression level of trpB in our experiment indicated a similar up-regulation, the expression levels of trpA did not change. As an additional strategy, we attempted to identify chlamydial genes involved in ADP/ATP exchange and energy source pathway reactions in the C. trachomatis genome. This analysis revealed six targets which may be involved in chlamydial persistence (a) two genes encoding proteins involved in the glycolysis pathway (pyk, yggV) (b), two genes (cydA, cydB) encoding proteins involved in the electron transport system, and (c) two genes encoding proteins involved in the production of tryptophan synthase subunits.