Human PBMCs were incubated with varying concentrations of prednisolone (10 nM, 100 nM, 10 μM or DMSO control) with LPS concentrations of 0.01 μg/mL, 1 μg/mL or vehicle. PBMCs were incubated with prednisolone for 30 min at 37°C. Cells were then stimulated with LPS for 2 hr at 37°C. Cell lysate was sent to Covance Genetis Lab (Seattle, WA) for analysis. Gene expression was analyzed in cell lysates with the NanoString nCounter system. Representative genes are used in this figure: transrepressed genes IFN-γ (a) and IFIT3 (b), transactivated gene PER1 (d) and MGEA5 as a negative control (c). Y-axis: Background correction (2) positive control correction and (3) housekeeping gene correction. The housekeeping genes were GAPDH, TUBB, GUSB, HPRT1, and PGK1. Positive control is undisclosed by nanostring. Data points represent n = 3. Error bars represent standard deviation.