NFATp and TB Infection Figure 5. Varying NFATp requirement for cytokine expression in different primary cell populations. A and B. TNF expression in BM- derived dendritic cells is not NFATp-dependent. Primary BM-DC were derived from WT and NFATp2/2 mice. After 7 days cells were stimulated with MTb lysates. At indicated time points the cells were lysed to prepare total RNA. (A) TNF mRNA was measured by real-time PCR. (B) TNF protein levels in the supernatants were measured by ELISA. C and D. TNF and IFN-c expression in murine primary T cells is NFATp-dependent. Purified primary total CD4+ and na¨ıve (CD4+CD62L+CD45RBhigh) T cells from uninfected WT and NFATp2/2 mice were stimulated immediately with anti-CD3/CD28 antibody and then left in culture in neutral conditions for another six days and re-stimulated with anti-CD3/CD28 antibodies. Aliquots of cells were PLoS ONE | www.plosone.org 7 July 2012 | Volume 7 | Issue 7 | e41427