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. 1997 May 13;94(10):5000–5005. doi: 10.1073/pnas.94.10.5000

Figure 3.

Figure 3

Cell surface expression of M2tag and M2-V27S oligomers in oocytes of X. laevis. Synthetic mRNAs encoding M2tag, M2-V27S, or a mixture of the two RNAs and using water as a control were microinjected (50 nl of RNA of 0.85 μg/μl for M2tag, 50 nl of 0.25 μg/μl for M2-V27S, and 50 nl of RNA containing 0.75 μg/μl M2tag mixed with 0.25 μg/μl M2-V27S) into oocytes of X. laevis. At 24 h after injection, oocytes were labeled with [35S]methionine for 18 h and surface-expressed M2 protein captured with M2-specific mAb (14C2). Oocytes were washed, homogenized, and detergent lysates prepared in the presence of 50 mM iodoacetamide as described. Antibody was recovered by adding protein A-Sepharose beads and immune complexes were analyzed by SDS/PAGE under nonreducing conditions.