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. Author manuscript; available in PMC: 2013 May 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2012 Mar 8;32(5):1246–1254. doi: 10.1161/ATVBAHA.111.241257

Figure 2. IL-8 induction is strongly regulated by HBEGF.

Figure 2

(A) HBEGF silencing decreased IL-8 induction by Ox-PAPC in HAECs. (B–C) Overexpression of HBEGF in HAECs (B) or the introduction of HBEGF in HEK293 cells (C) increased IL-8 expression induced by Ox-PAPC. Endogenous HBEGF level was undetectable in untransfected HEK293 cells. After 2 days following transfection of siRNA or HBEGF plasmid, cells were treated with Ox-PAPC (50ug/ml) for 4hrs and the IL-8 mRNA levels were determined by qRT-PCR. Western blotting and qRT-PCR were repeated at least three times and representative results shown. The silencing of target proteins was confirmed by qRT-PCR as shown in Suppl. Figure I. (D) Active HBEGF is increased in response to Ox-PAPC. HAECs were treated with Ox-PAPC (50ug/ml) for 0, 10, and 30min in duplicate, and the proteins in cell lysates were separated using SDS-PAGE. The formation of active HBEGF was determined by Western blotting. (E) Recombinant HBEGF significantly increased IL-8 in HAECs. HAECs were treated with HBEGF for 4 hrs and the levels of IL-8 were determined by qRT-PCR. The values were represented as mean + SD, *p<0.05 and ** p<0.01, N=3 for reactions.