Figure 4

Loss of WISP1 results in reduced expression of TGFβ1- and TNFα-induced IL-6 in phLFs.

phLFs were transfected with control (siCtrl) or WISP1-targeting (siWISP1) siRNAs and subsequently treated with TGFβ1 (2 ng/ml) or TNFα (10 ng/ml) for (A,B) 24 hours or (C-F) 24 and 48 hours. WISP1 (A) mRNA was analysed by RT-qPCR and (B) secretion was measured by ELISA. siRNA treatment against WISP1 results in significant loss of WISP1 at baseline and in the presence of TGFβ1 and TNFα treatment. IL-6 levels were analysed at 24 and 48 hours of 2 ng/ml TGFβ1 or 10 ng/ml TNFα treatment and (C,E) IL-6 mRNA was analysed by RT-qPCR and (D,F) IL-6 secretion was measured by ELISA. Loss of WISP1 results in loss of IL-6 induction even in the presence of TGFβ1 or TNFα treatment (n = 4; *^,#p < 0.05; **p < 0.01; ***^,###p < 0.001; 1-way ANOVA followed by Neuman-Keuls multiple comparison test).