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. 2009 Sep 28;106(40):17031–17036. doi: 10.1073/pnas.0905299106

Fig. 2.

Fig. 2.

Expression and secretion of IL-6 and IL-8 depend on IL-1α signaling. (A) HCA2 cells were either untreated or treated with bleomycin and allowed to recover for 6 d. Neutralizing antibodies or rIL-1ra were added for 24 h. CM were collected and analyzed for IL-6 (black bars) and IL-8 (gray bars) by ELISA. Cytokine levels were normalized to those secreted by untreated control cells. Neutralizing antibodies and recombinant proteins were added at the following final concentrations: IL-1α, 0.6 μg/ml; IL-1β, 0.8 μg/ml; rIL-1ra, 240 ng/ml. (B) Whole cell lysates were prepared from cells treated with the indicated neutralizing antibodies 7 d after bleomycin treatment and analyzed by Western blotting for IRAK1 and actin (control) protein levels. (C) Cells infected with shGFP or shIRAK1-expressing lentiviruses were treated with bleomycin and allowed to recover for 6 d. CM were collected over the next 24 h and analyzed for IL-6 by ELISA.