Invasive and Adhesive Properties of 32Dcl3 and BMC Expressing BCR/ABL Proteins
| Function . | Vector . | Wild-Type . | K1172R . | ΔSH3 . | P1013L . |
|---|---|---|---|---|---|
| 32Dcl3 | |||||
| Invasion chamber* | |||||
| Membrane | 1.9 ± 0.4 | 7.0 ± 0.4 | 2.0 ± 0.2 | 2.9 ± 0.2 | 3.7 ± 0.8 |
| Well | 30.7 ± 4.8 | 83.2 ± 6.0 | 31.2 ± 2.7 | 39.7 ± 1.9 | 52 ± 8.0 |
| Invasion of monolayer† | 135 ± 32 | 621 ± 113 | 119 ± 27 | 237 ± 19 | |
| Adhesion to stroma‡ | |||||
| Stroma MP− | 0.06 ± 0.03 | 1.04 ± 0.16 | 0.08 ± 0.02 | 0.44 ± 0.06 | |
| Stroma MP+ | 3.18 ± 0.53 | 0.16 ± 0.03 | 2.68 ± 0.37 | 0.40 ± 0.21 | |
| Adhesion to substrate§ | |||||
| Collagen IV | 17.0 ± 4.0 | 156 ± 24 | 16 ± 8 | 21 ± 13 | 50 ± 20 |
| Laminin | 26 ± 15 | 80 ± 8 | 23 ± 7 | 26 ± 12 | 37 ± 8 |
| Fibronectin | 121 ± 22 | 21 ± 5 | 127 ± 21 | 25 ± 9 | 38 ± 5 |
| Poly-L-lysine | <1 | <1 | <1 | <1 | <1 |
| BMC | |||||
| Invasion chamber∥ | |||||
| Membrane | 0.6 ± 0.3 | 3.2 ± 0.3 | 0.7 ± 0.2 | 1.1 ± 0.3 | |
| Well | 5.0 ± 2.0 | 19.0 ± 3.6 | 4.3 ± 2.1 | 7.7 ± 1.5 | |
| Adhesion to substrate§ | |||||
| Collagen IV | 4.6 ± 0.6 | 16.2 ± 0.9 | 5.2 ± 0.2 | 6.8 ± 0.4 | |
| Laminin | 6.4 ± 0.9 | 16.5 ± 1.9 | 6.1 ± 0.9 | 7.3 ± 0.6 | |
| Fibronectin | 14.5 ± 1.5 | 6.5 ± 0.7 | 14.0 ± 1.9 | 6.8 ± 0.5 | |
| Poly-L-lysine | 58.7 ± 7.1 | 60.0 ± 8.5 | 59.3 ± 3.2 | 58.3 ± 4.2 |
| Function . | Vector . | Wild-Type . | K1172R . | ΔSH3 . | P1013L . |
|---|---|---|---|---|---|
| 32Dcl3 | |||||
| Invasion chamber* | |||||
| Membrane | 1.9 ± 0.4 | 7.0 ± 0.4 | 2.0 ± 0.2 | 2.9 ± 0.2 | 3.7 ± 0.8 |
| Well | 30.7 ± 4.8 | 83.2 ± 6.0 | 31.2 ± 2.7 | 39.7 ± 1.9 | 52 ± 8.0 |
| Invasion of monolayer† | 135 ± 32 | 621 ± 113 | 119 ± 27 | 237 ± 19 | |
| Adhesion to stroma‡ | |||||
| Stroma MP− | 0.06 ± 0.03 | 1.04 ± 0.16 | 0.08 ± 0.02 | 0.44 ± 0.06 | |
| Stroma MP+ | 3.18 ± 0.53 | 0.16 ± 0.03 | 2.68 ± 0.37 | 0.40 ± 0.21 | |
| Adhesion to substrate§ | |||||
| Collagen IV | 17.0 ± 4.0 | 156 ± 24 | 16 ± 8 | 21 ± 13 | 50 ± 20 |
| Laminin | 26 ± 15 | 80 ± 8 | 23 ± 7 | 26 ± 12 | 37 ± 8 |
| Fibronectin | 121 ± 22 | 21 ± 5 | 127 ± 21 | 25 ± 9 | 38 ± 5 |
| Poly-L-lysine | <1 | <1 | <1 | <1 | <1 |
| BMC | |||||
| Invasion chamber∥ | |||||
| Membrane | 0.6 ± 0.3 | 3.2 ± 0.3 | 0.7 ± 0.2 | 1.1 ± 0.3 | |
| Well | 5.0 ± 2.0 | 19.0 ± 3.6 | 4.3 ± 2.1 | 7.7 ± 1.5 | |
| Adhesion to substrate§ | |||||
| Collagen IV | 4.6 ± 0.6 | 16.2 ± 0.9 | 5.2 ± 0.2 | 6.8 ± 0.4 | |
| Laminin | 6.4 ± 0.9 | 16.5 ± 1.9 | 6.1 ± 0.9 | 7.3 ± 0.6 | |
| Fibronectin | 14.5 ± 1.5 | 6.5 ± 0.7 | 14.0 ± 1.9 | 6.8 ± 0.5 | |
| Poly-L-lysine | 58.7 ± 7.1 | 60.0 ± 8.5 | 59.3 ± 3.2 | 58.3 ± 4.2 |
Results represent the mean ± SD from three independent experiments using individual 32Dcl3 clones or mixtures of BMC expressing the indicated BCR/ABL proteins.
Values are the number of cells (×103) on the matrix membrane (membrane) or collected at the bottom of the chamber (well) (mean ± SD, 3 experiments).
Number of clonogenic cells infiltrating stromal bone marrow monolayers (mean ± SD, 3 experiments).
Nonadherent and adherent cells (after trypsinization) were plated in MethoCult H4230 semisolid medium. Results are given as the ratio between the number of colony-forming cells in the adhering versus the nonadhering fraction (mean ± SD, 3 independent experiments) in the presence of recombinant murine IL-3 (10 U/mL).
Number (×104) of cells adherent to the indicated substrates (mean ± SD, 3 experiments).
∥After infection with the retroviral constructs, BMC were grown for 14 days in G418-containing medium in the presence of Kit-ligand, IL-3, and IL-6.