For soft agar assays, equal numbers of the indicated BJAB or Daudi cells (250, 500, 1000 or 2000 cells) were placed in soft agar containing DMEM, 20% FBS and 0.3% bacto agar (Difco, Franklin Lakes, NJ), and plates were placed at 37°C in a humid incubator with 5% CO2. To confirm cell counts, total cell protein assays (Bio-Rad) were performed on the cell dilutions used for plating. Macroscopic soft agar colonies were counted 14 days after plating.
Tumor studies were performed essentially as described previously (Yamamoto et al., 2000 (link); Gapuzan et al., 2002 (link)). 5 × 106 cells were injected subcutaneously into SCID mice (Taconic Farms, Germantown, NY). Once tumors appeared, mice were monitored 3× weekly and animals were sacrificed when tumors reached 2.25 mm2. All animal studies were performed in accordance with NIH guidelines and with approval of the Boston University Institutional Animal Care and Use Committee.
Tumor studies were performed essentially as described previously (Yamamoto et al., 2000 (link); Gapuzan et al., 2002 (link)). 5 × 106 cells were injected subcutaneously into SCID mice (Taconic Farms, Germantown, NY). Once tumors appeared, mice were monitored 3× weekly and animals were sacrificed when tumors reached 2.25 mm2. All animal studies were performed in accordance with NIH guidelines and with approval of the Boston University Institutional Animal Care and Use Committee.
