Fig. 3.

Quantification of CCT-alpha and CCT-zeta mRNA levels in healing adult wounds versus adult control assayed by real time RT-PCR. Real time RT-PCR was done to confirm patterns of expression of CCT-alpha and CCT-zeta, the two subunits for which full-length sequence was now available. The primers and Taqman probes were designed using Primer Express software (Applied Biosystems, Foster City, CA, USA) and are listed in Table 2. Forward and reverse primers were purchased from Integrated DNA Technologies (Coralville, IA, USA) and fluorocoupled Taqman probes were purchased from Applied Biosystems. The reverse transcriptase (RT) reaction (using reverse primer) and subsequent real-time PCR assays were performed as previously described (Kathju et al. 2006). Using the comparative critical cycle (Ct) method and using GAPDH as the endogenous control, the expression levels of the target genes were normalized and the relative abundance was calculated. Data were analyzed using the 7,900-HT SDS software version 2.1 provided by Applied Biosystems. Data are shown as mean ± SEM of six independent studies performed in duplicate. Statistical analysis was performed by Student’s t test (*p < 0.05). CCT-alpha displays a significant reduction in the adult wound tissue compared to unwounded control, while CCT-zeta does not