One ug RNA was used for first strand cDNA synthesis (Roche).
Fifteen ug of protein was loaded on a NuPAGE 4−12% Bis-Tris Gel (Invitrogen).
One ug of RNA was reverse-transcribed using the SuperScript II (Invitrogen); real-time PCR was performed as described [41].
One ug of RNA was used in each reaction to obtain the first-strand complementary DNA by reverse transcription using random primers (Promega).
Five ug mononucleosomes and naked genomic DNA control were labeled by biotin following the Affymetrix labeling kit protocol and hybridized in parallel to tiling arrays.
One ug of RNA was reverse-transcribed to cDNA using oligo dT) primers with RT-PCR kit according to the manufacturer's instructions (Invitrogen Corp., Carlsbad, CA, USA).
Fifty ug of each sample was subjected to fluorescence labeling using 2 dyes (Cy3 for plasma and Cy5 for lymph) and to 2D-DIGE preparative electrophoresis (Applied BIOMICS Inc. facilities, Hayward, CA).
Ten ug of genomic DNA from frozen spleen sections and bone marrow isolated at harvest was digested with EcoR1 and Sal1, run out on 0.8% agarose gel, transferred to Hybond-N+ (GE Healthcare, Piscataway, NJ, USA) and hybridized with a radioactive probe from the IRES gene in MIG-R1.
Ten ug of total RNA from each sample was hybridized to Sera-mag oligo (dT) beads (Thermo Scientific) for mRNA purification.
