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. 2014 Jun 13;193(2):889–900. doi: 10.4049/jimmunol.1303389

FIGURE 2.

FIGURE 2.

sCTLA-4 is secreted and expressed as a glycosylated monomer. (A) Human Tm-CTLA-4 protein is detected as a dimer under nonreducing conditions (NR) and as a monomer under reducing conditions (R) in lysates from HeLa cells transfected with human Tm-CTLA-4 cDNA. (B) Lysates from HeLa cells transfected with human sCTLA-4 cDNA contain sCTLA-4 protein that migrates as a monomer in both nonreducing and reducing conditions. (C) Under reducing conditions, sCTLA-4 protein secreted into the culture medium of HeLa cells transfected with sCTLA-4 cDNA migrates more slowly than sCTLA-4 present in cell lysates. (D) N-glycan addition contributes to the molecular masses of Tm-CTLA-4 and sCTLA-4. Following culture with or without tunicamycin, detergent lysates from HeLa cells transfected with Tm-CTLA-4 or sCTLA were analyzed by Western blot under reducing conditions using C19 polyclonal Ab or 4B8 mAb, respectively. Arrows indicate the protein species lacking N-linked glycans. All experiments are representative of at least three independent observations.