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. Author manuscript; available in PMC: 2013 Oct 12.

Published in final edited form as: J Mol Biol. 2012 Jun 25;423(1):47–62. doi: 10.1016/j.jmb.2012.06.023

Fig. 3 — (A) Schematic of constructs for Furin and PC1. (B) Normalized protease activity assayed in conditioned media (CM) from Cos-7 cells transfected with 2 μg of DNA (C) Western blot analysis of CM from cells expressing secreted reporter constructs (top panel; SEC), and ER fractions from cells expressing KDEL-tagged reporters probed using mAB-M2. Molecular weight of each species is indicated by the arrowheads; Unprocessed furin, 78kD; Processed furin, 69kD; Unprocessed PC1, 66kD; Processed PC1 57kD. (D) pH-dependent activation of KDEL-tagged reporters measured after incubating ER membrane fractions at designated pH²⁹. Maximal activity was estimated by trypsinizing membrane fractions for 1 hr and inhibiting trypsin by soybean trypsin inhibitor prior to the protease assay.

Fig. 3 — (A) Schematic of constructs for Furin and PC1. (B) Normalized protease activity assayed in conditioned media (CM) from Cos-7 cells transfected with 2 μg of DNA (C) Western blot analysis of CM from cells expressing secreted reporter constructs (top panel; SEC), and ER fractions from cells expressing KDEL-tagged reporters probed using mAB-M2. Molecular weight of each species is indicated by the arrowheads; Unprocessed furin, 78kD; Processed furin, 69kD; Unprocessed PC1, 66kD; Processed PC1 57kD. (D) pH-dependent activation of KDEL-tagged reporters measured after incubating ER membrane fractions at designated pH²⁹. Maximal activity was estimated by trypsinizing membrane fractions for 1 hr and inhibiting trypsin by soybean trypsin inhibitor prior to the protease assay.