Involvement of signaling components and change of HLA-I, CD40 and CD54 expression upon combined cytokine treatment with IL-4 and TNF α in RCC cell lines. Hal31RC was treated with either IL-4 or TNFα alone and with the combination of the two cytokines. A) Intracellular pSTAT6 as well as NFκB (pS529) at 30 min post stimulation were analyzed by flow cytometry. Gray filled graph represents constitutive expression and black bold line represents cytokine-induced expression of either signaling component. Numbers on gates are percentage of positive cells compared to medium control. Representative data of at least 5 different experiments are shown. B) HLA class I, CD40, CD54 expression 72 hrs post cytokine treatment were analyzed by flow cytometry. Data are presented as fold increase when compared to constitutive expression of these molecules on untreated Hal31RC. Data are mean ± SEM of 9 different experiments.