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. 2010 Sep 15;21(18):3093–3105. doi: 10.1091/mbc.E10-04-0356

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© 2010 by The American Society for Cell Biology

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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Figure 5. — Combined knockdown of PDI and ERp57 substantially impairs protein folding. HepG2 cells constitutively depleting ERp57 by shRNAmir were subjected to knockdown of PDI by siRNA for 6 d. HepG2 cells transfected with control siRNA were prepared in parallel. Pulse-chase experiments were conducted and analyzed as described in Figure 3. (A) Kinetics of disulfide formation. (B) Secretion kinetics. Amount secreted was quantified as a percentage of the total signal recovered at the 10 min chase time point. Black and gray bars represent control and knockdown conditions, respectively. (C) Quantification of oxidative folding. Solid lines, control conditions; dashed lines, knockdown conditions. (D) ER to Golgi transport kinetics. (E) Quantification of the gels in panel D. Error bars represent the average of three independent experiments ± one SD, except in the case of TF which was examined in a single experiment. Asterisks indicate quantifications were not included beyond 30 min of chase due to substrate degradation.