Fig. S7

T-LBL Cells Overexpressing bcl-2 and Myc are smaller, resistant to apoptosis, and from Aggregates at Low Cell Density and under Hypoxic Conditions, without active proliferation. (A-L) Brightfield images of T-LBL cells on ZKS stromal cells after 7 days of culture following serial dilution and plating (A-E and G-K), or after 8 days in hypoxic conditions (F and L). All FACS-sorted tumor cells were either from 7 Myc;Cre;bcl-2 T-LBL (top row) or Myc;Cre T-LBL (bottom row) transgenic fish. (M) Ratio of aggregates over free cells for tumor cell cultures when diluted 1:1 (black bars), 1:2 (gray bars), 1:4 (light gray bars), 1:8 (white bars), and 1:16 (black hatched bars), and grown for 7 days: Myc;Cre T-LBL (n=3; P=0.017 for 1:1 dilution; P=0.00185 for 1:2 dilution), Myc;Cre;bcl-2 T-LBL (n=4), Myc;Cre;bcl-2 T-ALL (n=4; P=0.0004 for 1:1 dilution; P=0.0021 for 1:2 dilution; P=0.017 for 1:4 dilution ), or Myc;Cre;bcl-2;Myr-Akt2 T-ALL (n=5; P=0.0018 for 1:1 dilution; P=0.0137 for 1:2 dilution; P=0.028 for 1:4 dilution ) transgenic fish. (N) Average diameters of cells under hypoxic (gray) and normal (black) culture conditions for Myc;Cre;bcl-2 vs. Myc;Cre tumor cells (mean ± SD μm diameter of cell under normal conditions: 1.79 ± 0.59 vs. 3.33 ± 1.50; n=110 for each group, P<0.0001; mean ± SD μm diameter of cell under hypoxic conditions: 1.62 ± 0.55 vs. 3.30 ± 1.46; n=110 for each group, P<0.0001;). (O) Fold change in viable lymphoma or leukemic cells between 1 and 7 days postculture on ZKS stroma cells from Myc;Cre T-LBL (dark gray bar, n=3), Myc;Cre;bcl-2 T-LBL (light gray bar, n=4), Myc;Cre;bcl-2;Akt2 (white bar, n=5), and Myc;Cre;bcl-2 T-ALL (black bar, n=4) transgenic fish. (P) Percentage of Annexin V-positive cells from whole kidney marrow (WKM; black bar), Myc;Cre T-LBL (dark gray bar, n=3), Myc;Cre;bcl-2 T-LBL (light gray, n=4), Myc;Cre;bcl-2;Akt2 (white bar, n=5), and Myc;Cre;bcl-2 T-ALL (black bar, n=4) cells after 7 days in culture. Data are reported as means ± SD. Scale bar for panels A-L = 40 μm.