TgCtwh3 and TgCtwh6 induced different levels of cytokines. (A and B) BMMφs were infected with either RH, or PRU, or TgCtwh3 or TgCtwh6 tachyzoites, and 24 hr later supernatants were harvested and cytokine production of IL-12p40 (A) and TNF-α (B) were determined by ELISA. (C and D) RNA extracted from RH-, PRU-, TgCtwh3- or TgCtwh6-infected BMMφs, and mRNA was analyzed by real time-PCR using corresponding specific primers. Uninfected Mφ used as negative control was run in parallel. Housekeeping gene GAPDH allowed normalization of the expression of the interested genes. Relative expressions of IL-12p40 (C) and TNF-α (D) were displayed. Bars represent means ± SD from three independent experiments (#p>0.05, *p<0.05, **p<0.01, ***p<0.001, parasites infected cells vs control).