Gelelectrophoresis and melting curve analysis confirmed the presence of the expected PCR products only, and the absence of unwanted non-specific products (data not shown). Non-inoculated RHE failed to show evidence of gene Sotrastaurin expression (data not shown), confirming that each primer pair was specific for its corresponding C. albicans gene. Using

the optimized real-time PCR assays, we found that HWP1 and all ALS, SAP, LIP and PLB genes were expressed at all time points during biofilm growth in all model systems tested (and also in the start cultures), as evidenced from a detectable Ct value (Ct < 35; data not shown). Expression levels of ALS genes and HWP1 in biofilms The expression levels (expression in biofilms, relative to expression in start cultures) of ALS genes and HWP1 in biofilms at selected time points in the various model systems are shown in Additional file 1. ALS1-5 were overexpressed in biofilms grown in all model systems at several time points or during the entire time course. Furthermore, HWP1 and ALS6 were overexpressed

in all model systems except in the MTP and RHE, respectively. ALS9 was only overexpressed in biofilms grown in the CDC reactor, but the fold upregulations were not particularly high. The fold expressions were model-dependent for most of the genes tested. Overexpression of ALS3 and HWP1 were more pronounced in biofilms grown in the in vivo model, while the expression levels of ALS6 were higher in the two in vitro models. Furthermore, the fold upregulations of ALS4 were more pronounced in biofilms grown in the in vivo and RHE models, while those of ALS1, ALS2 and ALS5 were higher in GSK2118436 the two in vitro models and in the in vivo model. Expression levels of SAP genes in biofilms The expression levels of SAP genes in biofilms at selected time points in the various model systems are shown in Additional file 2. All SAP genes (except SAP3) were upregulated in biofilms grown in

all model systems at one or more time points. The expression Niclosamide levels of SAP3 were rather erratic, and this gene was not considerably upregulated in any of the model systems tested. For most of the SAP genes model-dependent expression levels were observed. In in vitro grown biofilms, SAP1, SAP2, SAP4 and SAP6 were highly upregulated, and the fold expression of SAP2, SAP4 and SAP6 were also high in the vivo model. Furthermore, SAP5 was highly upregulated in biofilms grown in the in vivo and RHE models. Only for SAP9 and SAP10 similar gene expression levels were observed in all model systems, although these genes were not expressed at a high level in biofilms. Expression levels of PLB genes in biofilms The expression levels of PLB genes in biofilms at selected time points in the various model systems are given in Additional file 3. Overall, PLB genes were not considerably upregulated in biofilms, and only model-dependent differences in gene expression levels were observed.