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. 2010 Jan 25;285(14):10376–10384. doi: 10.1074/jbc.M109.055947

FIGURE 4.

FIGURE 4.

Identification of an arginine-based ER retention signal and key residues required for ER retention. A, surface and total expression of TacADAM10 arginine mutants expressed in COS cells. B, surface/total expression ratios of TacADAM10 mutants were measured and normalized to TacADAM10 surface/total ratios (*, p < 0.001 versus TacADAM10; #, p < 0.001 RRR to AAA and RRR to AAR versus RRR to RAR; n = 14–27 cells per condition). Scale bar, 20 μm. C, Western blot analysis of Tac, TacADAM10, and point mutants. D, deglycosylation analysis reveals that RRR to ARR and RRR to RRA are sensitive to EndoH deglycosylation, indicating that they are retained in the ER. In contrast, higher molecular weight bands corresponding to RRR to AAA, RRR to AAR, and RRR to RAR mutants are resistant to Endo-H indicating that they leave the ER and enter the Golgi. Open circles, EndoH-resistant bands; asterisks, EndoH-sensitive bands.