American Society of Hematology

Figure 2

$Figure 2. Wnt3a has no direct effect on growth of MM cell lines and primary MM cells. Cells from indicated MM cell lines were cultured in serum-free medium in the absence and presence of rWnt3a for the indicated times. Cell proliferation was measured by MTT assay as described in “Wnt3a does not promote MM growth.” (A) Protein isolated from H929/W3A or H929/EV was subjected to Western blotting analysis using anti-HA antibody to confirm Wnt3a expression (top) or using anti–β-catenin antibody (second panel from top). The same fractions were also blotted with antitubulin antibody as a control for protein loading (bottom). The proteins isolated from H929/W3a or H929/EV was subject to E-cadherin pull-down assay to measure the uncomplexed β-catenin (second panel from bottom). (B) H929/W3A and H929/EV cells were seeded in RPMI supplemented with 10% FBS for the indicated time, and proliferation was measured by MTT assay. (C) MM cell lines H929, INA6, and OPM-2 were incubated for 48 hours in the absence and presence of rWnts (100 ng/mL) and then subjected to MTT assay. (D) Primary CD138-selected plasma cells from 2 patients with MM (Pt no. 1 and Pt no. 2) were cultured in serum-free medium in the presence of Wnt3a-CM or Cont-CM or with rWnt3a protein for the indicated time. Cell proliferation was measured by MTT assay. Results are expressed as means plus or minus SEM.$

Wnt3a has no direct effect on growth of MM cell lines and primary MM cells. Cells from indicated MM cell lines were cultured in serum-free medium in the absence and presence of rWnt3a for the indicated times. Cell proliferation was measured by MTT assay as described in “Wnt3a does not promote MM growth.” (A) Protein isolated from H929/W3A or H929/EV was subjected to Western blotting analysis using anti-HA antibody to confirm Wnt3a expression (top) or using anti–β-catenin antibody (second panel from top). The same fractions were also blotted with antitubulin antibody as a control for protein loading (bottom). The proteins isolated from H929/W3a or H929/EV was subject to E-cadherin pull-down assay to measure the uncomplexed β-catenin (second panel from bottom). (B) H929/W3A and H929/EV cells were seeded in RPMI supplemented with 10% FBS for the indicated time, and proliferation was measured by MTT assay. (C) MM cell lines H929, INA6, and OPM-2 were incubated for 48 hours in the absence and presence of rWnts (100 ng/mL) and then subjected to MTT assay. (D) Primary CD138-selected plasma cells from 2 patients with MM (Pt no. 1 and Pt no. 2) were cultured in serum-free medium in the presence of Wnt3a-CM or Cont-CM or with rWnt3a protein for the indicated time. Cell proliferation was measured by MTT assay. Results are expressed as means plus or minus SEM.

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