Fig. 6: Syndecan-1 controls the spatiotemporal dynamics of bcCML in vivo.

a Schematic illustrates the experimental approach used to assess the impact of Sdc1 loss on the dynamics of bcCML in vivo. bcCML cells infected with shCTRL or shSdc1were transplanted into irradiated dsRed NOD SCID recipient mice. The calvarium of recipient mice was live imaged 10–15 days post-transplantation, after which the femurs and spleens were analyzed by flow cytometry. b Confirmation of Sdc1 knockdown in bcCML cells infected with shSdc1 as quantified as a reduction in Sdc1 MFI by flow cytometry. Significance from shCTRL within a location (n = 2 mice per group). c Representative live animal imaging of dsRed recipient mice transplanted with shCTRL or shSdc1 bcCML; green = shRNA vector, blue = VE-Cadherin, magenta = Actin dsRed, white arrowhead shows cells within the vasculature, red arrowhead shows cells outside of vasculature within the marrow, yellow arrowhead shows cell clusters (See also Supplementary Movies 2 and 3). d Quantification of GFP⁺ bcCML burden from live animal imaging of bone marrow (xyz confocal stacks, n = 2 animals per group, open and closed symbols denote separate animals, n = 10 fields of view per group). e Quantification of GFP⁺ bcCML residing within vasculature determined by live animal imaging (xyz confocal stacks, n = 2 animals per group, open and closed symbols denote separate animals, n = 10 fields of view per group). f The average velocity of GFP⁺ bcCML determined by measuring in vivo cell migration in real time (xyt confocal stacks, n = 2 animals per group, ≥3 fields of view per mouse, shCTRL n = 25 cells, shSdc1 n = 15 cell). g Representative flow cytometry plots show the distribution of bcCML in the femur and spleen of mice used in live animal imaging studies in (b–f). Percentages of shRNA-infected Sdc1+ and Sdc1− bcCML cells are indicated. h Quantification of tumor burden in femur and spleen of mice used in the live animal imaging studies (n = 2 animals per group). Two-tailed unpaired Student’s t-tests were used to determine statistical significance. Source data are provided as a Source Data File.