Figure S1 from A Subpopulation of Stromal Cells Controls Cancer Cell Homing to the Bone Marrow

Supplementary-Figure 1 Tumor cell infiltration of other organs after PTH/ZA treatment (A) The combined treatment with PTH/ZA does not affect the homing of MDA tumor cells to other organs; n=14/15 in 2 experiments. Evaluation of blood vessel numbers and binding of cancer cells to vascular cells after PTH/ZA treatment (B) PTH/ZA did not affect blood vessel numbers in the treated mice. Proximal tibiae from 4 mice/group were evaluated and the number of blood vessels adjusted to the evaluated area. (C) Binding of cancer cells to immune cells (CD45+) or vascular cells (CD31 or PLVAP) was not affected by treatment with PTH/ZA, n=13/11. Bone marrow from control and treated mice that received cancer cells 24 hours prior to bone marrow isolation was subjected to addition of magnetic beads precoated with CD45 antibody and carefully removed. Addition of either CD31 or PLVAP antibody coated beads followed and beads were used for DNA isolation and evaluation of cancer cell presence. The amount of cancer cell specific DNA was corrected to murine -actin. PTH/ZA treatment does not affect colony forming units of hematopoietic populations (D) There were no changes in various progenitor colonies when bone marrow from treated mice was cultured in vitro for 2 weeks in specialized media, stained with crystal violet and counted; n=6/6/6/6 in 3 experiments. CFU-GEMM is shown in Fig. 1C. Examples of the various colonies are shown below the graphs (light microscopy, 40x).