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. 2013 Feb;27(2):568–580. doi: 10.1096/fj.12-209445

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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/us/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — The β3 Ig domain binds β3 and β1. A–C) HEK293 cells were cotransfected with plasmids expressing β3-Ig-6His and β3-wt-EGFP(A); β1-wt (B), or β3-ΔECD-EGFP (C). Lysates were precipitated with Talon (anti-His) beads and blotted with antibodies as indicated. D) Control lanes showing cells transfected with only β3-Ig-6His together with untransfected cells. Lysates were precipitated with Talon (anti-His) beads and blotted with antibodies as indicated. FT, flow through; W, wash; E, elution fraction. E) PNGase F digestion reduces the higher-molecular-mass doublet characteristic of β3-Ig-6His protein to a single band of 15–16 kDa. Mwt, molecular weight (mass). Asterisk indicates the added PNGase F enzyme.