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Figure 6.
Localization of sclerostin to the surface of cells. The top
section of panel A shows a Western blot (with an antibody to
sclerostin) of 24-h supernatants obtained from MC3T3-E1 cells
transfected with wild type human sclerostin. The transfections
and treatments were as follows: 1, empty vector; 2, sclerostin;
3, empty vector with heparin added to a final concentration of
500 μg/ml; 4-12, sclerostin with heparin added to final
concentrations of 500, 250, 100, 50, 25, 12.5, 1, 0.5, and 0.25
μg/ml, respectively. The bottom section of panel A shows the
Western blot of samples generated from corresponding wells
incubated with a lysis buffer to determine the total amount of
sclerostin being produced. The top section of panel B shows the
Western blot (with an antibody to sclerostin) obtained for
samples of 24-h supernatants from MC3T3-E1 cells transfected
with vectors encoding wild type or mutant sclerostin. The
transfections were as follows: 1, empty vector; 2, wild type
sclerostin; 3, sclerostin R114A, R116A, and R119A; 4, sclerostin
K134A and R136A; 5, sclerostin R97A, K99A, and R102A; 6,
sclerostin K142A, K144A, and R145A. The bottom section of panel
B shows the Western blot obtained for equivalent samples of
total cell lysates.
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