An siRNA was constructed (Ambion Inc., www.ambion.com) to target a sequence unique to the NFAT5 transcript: 5′-CAACATGCCTGGAATTCAA-3′ (nt 335 to 353) [31]. As described, a control for non-specific siRNA effects, we used an siRNA targeting the green fluorescent protein (GFP), 5′- GGCTACGTCCAGGAGCGCACC-3′. MDM were transfected in 6-well plates using 1 µM of the indicated siRNA in siPORT NeoFX transfection reagent (Ambion Inc., www.ambion.com), prepared as recommended by the manufacturer, in a final volume of 750 µl in Macrophage-SFM medium plus 5% heat-inactivated human AB serum. The cultures were left at 37°C overnight after which cells were washed and incubated in fresh medium. MDM were transfected two times for efficient knock down of NFAT5 expression before infection experiments were performed [31].