Steady-state p35 mRNA induced in monocytes is approximately 0.2 kb shorter than constitutive or enhanced mRNA in EBV-transformed lymphoblastoid cells (Northern analysis). Polyadenylated RNA (2 × 106 cell equivalents/lane) was isolated from the EBV-transformed cell line VDS that was either uninduced (lane 1) or stimulated with PMA (10 ng/mL) and A23187 (25 ng/mL) for 24 hours (lane 2), from monocytes primed with IFN-γ (100 ng/mL for 16 hours), followed by LPS (1 μg/mL) for 2 hours (lane 3), or from monocytes stimulated with LPS alone (lane 4). RNA was resolved by electrophoresis on a 1.5% agarose gel and blotted to a nylon filter. The blot was probed with a p35 cDNA probe as described in the Materials and Methods. RNA size was estimated using an RNA ladder (Life Technologies).
