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. 2013 Sep 11;8(9):e74094. doi: 10.1371/journal.pone.0074094

Figure 2. HTRA1 cleaves TGF-β receptors.

Figure 2

(A) Cleavage of TGF-β receptors by HTRA1 in vitro. Equal amounts of purified TGF-β receptors were incubated with increasing amount of purified HTRA1 with or without CPII, an HTRA1 agonist. The inactive mutant, S328A, was also incubated with the receptors in the presence or absence of CPII. Protein digestions were analyzed by western blot. UD = undigested proteins. (B) HTRA1 cleavage of TGF-β receptors from the cell surface. HeLa cells were transfected with either a control or HTRA1 plasmid in addition to plasmids encoding either TβRIII, TβRII or TβRI. Membrane proteins were isolated and analyzed by western blot for cleavage of membrane bound receptors. Cadherin was used as a loading control. Overexpression of His-tagged HTRA1 was confirmed with an anti-His antibody. (C) mRNA levels of the TGF-β receptors after HeLa cells were transfected with either a control or HTRA1 plasmid.