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. Author manuscript; available in PMC: 2009 Dec 10.
Published in final edited form as: Exp Cell Res. 2008 Oct 7;314(20):3724–3736. doi: 10.1016/j.yexcr.2008.09.019

Fig. 4. Phospho-JNK levels are not affected by T. gondii infection in wild type mouse embryonic fibroblasts and p65−/− cells following TNFα exposure.

Fig. 4

Fig. 4

(A) A representative immunoblot of uninfected and T. gondii infected wild type mouse embryonic fibroblasts following exposure to TNFα and harvested at the time points indicated. JNK-phosphorylation (PJNK) occurs within 15 minutes post-UV and is transient with a pattern similar to that seen in HeLa cells. This response is fundamentally identical with regard to the extent of JNK phosphorylation and its kinetics in cells infected with T. gondii. Total JNK (JNK) and calnexin serve as loading controls. (B) Densitometric analysis representing the level of phospho-p54 JNK relative to total JNK in four independent experiments depicts the range of values inherent in such experiments. Relative phospho-p54 levels in uninfected cells (circle) versus infected cells (diamond) confirm an absence of inhibition of mean phospho-p54 JNK accumulation (horizontal line) in parasite infected cells relative (diamond) to uninfected cells (circle). Densitometric analysis was performed as described in the Materials and methods (C) Statistical analysis using two way ANOVA indicate a modest effect for time post-treatment but not for infection. Time and infection do not exhibit significant interaction.. The identical experiment to that performed with wild type MEF was replicated in p65−/− fibroblasts. Results indicate the absence of any significant inhibition in relative phospho-p54JNK levels. (E) Densitometric analysis of six independent experiments confirms the broad range of responses and the lack of apparent differences in the mean levels of phospho-p54 JNK comparing uninfected (circle) and infected (diamond) p65−/− cells. (F) Two-way ANOVA analysis reveals a highly significant effect for time post-UV but not infection. Significance: p values of 0.01 and lower are considered highly significant (***), p values between 0.01 and 0.05 are considered significant (**) while those above 0.05 are not significant (ns).