Figure 1.

Concentration of CNX and not ERp57 in the juxtanuclear ERQC upon proteasomal inhibition. Double-label immunofluorescence was done on fixed and permeabilized NIH 3T3 cells (A–L) or HeLa cells (M–R), preincubated for 3 h in the absence or in the presence of 10 μM Lac as indicated. Rabbit polyclonal anti-CNX was used as primary antibody with Cy3-conjugated goat anti-rabbit IgG as the secondary in A–F and M–R. Rabbit polyclonal anti-ERp57 with Cy3-conjugated secondary was used in G–L. Double labeling was done with mouse monoclonal anti-β-COP in A–L or with mouse monoclonal anti-ERp57 in M-R with FITC-conjugated goat anti-mouse IgG. Each colored panel is a merged image corresponding to the red and green channels in the panels to their left and includes 4,6-diamidino-2-phenylindole staining of nuclei. Inserted in F is an enlargement of the same image. Bars, 10 μm.