A. Data are Affymetrix signals for CCR1 (probe set 205098_at) and CCR2 (probe set 206978_at) in LP1, XG-19 and XG-6 HMCLs. Membrane expression of CCR1 or CCR2 was evaluated by flow cytometry using PE-conjugated anti-CCR1 or anti-CCR2 MoAbs or isotype-related PE-conjugated control MoAbs. The numbers in the panels are the RFI of the PE-conjugated anti-CCR1 or anti-CCR2 MoAbs compared to the PE-conjugated control MoAbs.
B. The chemoattractant activity of osteoclasts to myeloma cells was assayed using XG-19, XG-6 and LP1 HMCLs. Data are the fraction of MMC in the upper chamber of the transwell that could migrate to the lower chamber. Results are the mean values ± SD of 3 experiments. * indicates a significant difference of MMC migration in MMC/osteoclast cocultures compared to control using a paired Student’s t test (P ≤ .05). ** indicates a significant difference of MMC migration in MMC/osteoclast cocultures with anti-CCR1 and/or anti-CCR2 MoAb compared to MMC/osteoclast cocultures using a paired Student’s t test (P ≤ .05).
C. The expression of CCR1 and CCR2 by CD138+ primary myeloma cells of patients was evaluated by flow cytometry using FITC-conjugated anti-CD138 MoAb and PE-conjugated anti-CCR1 or anti-CCR2 MoAbs labelling. Isotype matched FITC-conjugated or PE-conjugated MoAbs recognizing no human antigens were used as control MoAb. The numbers in the panels are the RFI of the PE-conjugated anti-CCR1 or anti-CCR2 MoAbs compared to the PE-conjugated control MoAbs.
D. The chemoattractant activity of osteoclasts to myeloma cells was assayed using primary myeloma cells. Data are the fraction of primary MMC in the upper chamber of the transwell that could migrate to the lower chamber.