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Figure 2


Fig. 2. Depletion of Sec13 causes a loss of peripheral ERES labelling, and an accumulation of COPII and cargo in the juxtanuclear region of cells. (A) Cells depleted of Sec13 for 72 hours using one or both of the siRNA duplexes (-1 or -2) were fixed and immunolabelled for Sec31A, Sec24C, GM130, Sec16 or ERGIC-53 as indicated. (B) Cells transfected with siRNA duplexes targeting both Sec31A and Sec31B were immunolabelled for Sec16 and ERGIC-53. Note the juxtanuclear accumulation of COPII (Sec16) and cargo (ERGIC-53) in both cases (arrows). (C) Lysates of cells depleted of Sec13 were immunoblotted with antibodies specific for ERGIC-53 and {alpha}-tubulin. Molecular-mass markers are shown in kDa. (D) Localization of Sec16A and ERGIC-53 at sub-saturating pixel intensities in lamin-A/C- or Sec13-suppressed cells. (E) Suppression of Sec13 expression causes a reduction in COPI labelling of both Golgi and peripheral punctae compared with lamin-A/C-suppressed controls. Sec31A labelling is used to show effective suppression of Sec13/31 expression. Scale bars: 10 µm.