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. 2009 Nov 16;285(2):914–922. doi: 10.1074/jbc.M109.010892

FIGURE 1.

FIGURE 1.

The level of wild type and mutant S100A4 proteins in transfected Rama 37 cell clones and cell pools. Cells were transfected with expression vectors containing S100A4 wild type or mutant 72, 72.78, or 16.72.78 cDNAs as described under “Experimental Procedures.” Extracts of cloned cell lines (A and B) or pooled transfectants (C and D) were separated by SDS-PAGE and blotted onto polyvinylidene difluoride membranes. Membranes were incubated with either a polyclonal S100A4 antibody (A and C) or anti-actin serum (B and D) as a loading control. Bound antibodies were detected by chemiluminescence with horseradish peroxidase-conjugated secondary antibody as described previously (28, 30, 31). The intensities of the bands of S100A4 protein corrected for actin levels in cloned cell lines, Rama 37, Rama 37 + vector, Rama 37 + S100A4 mutant 72, Rama 37 + S100A4 mutant 72.78, and Rama 37 + S100A4 mutant 16.72.78 were 16, 10, 95, 90, and 88% that of wild type and, for cell pools, 10, 5, 99, 110, and 91% that of wild type.