(a) The expression of mRNA coding for TF and effector molecules was measured by qPCR at various time points following the activation of naive CD4 T cells in the presence of Th1, Th2 or Th17 polarizing cytokines. Results are mean ±SEM fold change as compared to naive CD4 T cells from three to six independent experiments. Th1 cells expressed higher levels of TNF, GZMK, CX3CR1 than Th2 and Th17 cells, whereas ASCL2 and ZEB2 were expressed at high levels by more than one Th subset. (b) The expression of Runx3, T-bet, Eomes and ThPOK was measured by flow cytometry after 14 days of activation of naive CD4 T cells in the presence of Th1 polarizing cytokines. Results are median ±IC of MFI of perforin (left panel) and normalized MFI of transcription factors expression by perforin+ cells as compared to perforin- cells from four to six experiments. **:p<0.01, ***:p<0.001. Perforin+ cells expressed higher levels of Runx3, T-bet, Eomes and ThPOK than perforin- cells.