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. 2001 May 28;153(5):1011–1022. doi: 10.1083/jcb.153.5.1011

Figure 4.

Figure 4

GADD34 inhibits CHOP activation in the UPR. (A) FACS® analysis of CHO cells expressing the CHOP::GFP reporter and transfected with CD2-expressing plasmids that coexpress the indicated derivatives of mouse GADD34, the A1 GSE, or no additional protein. The cells were treated with tunicamycin (2 μg/ml, 16 h) to activate CHOP::GFP. Note the presence of CD2-positive and GFP-negative cells (rectangle) in the pools transfected with A1, wild-type GADD34, and GADD34ΔN. (B) Immunostaining of mouse embryonic fibroblast cells transfected with expression vectors encoding FLAG-tagged A1 or GADD34, and the indicated deletion mutants of GADD34. The cells were treated with tunicamycin (2 μg/ml, 8 h) and immunostained with an antibody to FLAG that detects the recombinant proteins and an antiserum to CHOP that detects endogenous CHOP. The karyophilic dye H33258 stains the nuclei of all the cells in the field. The arrows mark the cells expressing the recombinant GADD34 proteins. (C) High magnification (60×) view of transfected cells expressing the full-length mouse GADD34 or the ΔN deletion. The recombinant proteins are revealed by immunostaining of the FLAG epitope tag.