Real Time PCR analysis of T helper cells

Murine lymphocytes T helper cells were obtained from spleen of C57Bl/6 mice and miR-155 deficient mice strains that have been immunised with MOG peptide 35-55 in complete Freunds adiuvant 12 days prior. Subsequently cells were stimulated for 72 hours with MOG peptide 35-55 or left unstimulated followed by T helper cell sorting using magnetic beads and RNA isolation. qPCR gene expression profiling. Cells from three individual mice of the same genotype were pooled and treated separately as indicated in the summary. Equal amount total RNA from each mice was pooled prior to gene expression analysis.