Fig. 4

Validation by RT-PCR of superior KLF2, PPARγ, ARNTL, Lck, ZAP-70, and PTPN13 mRNA expression in CCR4⁺CCR6⁺ Th17 versus CXCR3⁺CCR6⁻ Th1 cells. Total RNA was extracted from Th17 and Th1 subsets isolated and stimulated via CD3/CD28 for 3 days as described in Fig. 1. Expression of KLF2, PPARγ, ARNTL, PTPN13, Lck, and ZAP-70 mRNA was quantified by SYBR green real time RT-PCR. Quantification was performed relative to a standard curve generated based on cDNA specific for each transcript. The expression of each gene was normalized to the 28S rRNA internal control (28S rRNA) and expressed as fgs RNA of a target gene per 1 ng rRNA28S. Depicted are results obtained with matched Th17 versus Th1 subsets isolated from n = 4 different HIV-uninfected individuals. Paired t test values are indicated on the graphs. Fold change (FC) expression values in Th17 versus Th1 are included in the graphs