Article
Extended Data Fig. 5 | Multicolour quantitative deep-tissue confocal
imaging of complete femoral sections from MFG (Mds1GFP/+Flt3Cre) mice.
a, Identification of C-KIT+GFP+MFG-HSCs using multicolour quantitative deep-
tissue confocal imaging of full bone femoral sections. Pictures are 10-μm xy
projections of one area of interest. n = 3 mice. The experiment was performed
three times with similar results. b, Example of one full-bone femoral section
with colour-coded visualization of HSCs based on their distance from bone.
Yellow squares represent individual HSCs in proximity to cortical or trabecular
bone, whereas green dots represent individual HSCs located more than 10 μm
away. The picture represents data from an individual mouse. The experiment
was performed three times with similar results (d). c, Example of full-bone
femoral section (only Col.1 and DAPI staining are shown). The experiment was
performed three times with similar results. d, Colour-coded visualization of
HSCs based on their distance to bone. Yellow squares represent individual
HSCs in proximity to cortical or trabecular bone, whereas green dots represent
individual HSCs located more than 10 μm away. This picture represents an
independent mouse from b. The experiment was performed three times with
similar results. e, Quantification of absolute number and anatomical location
of C-KIT+GFP+MFG-HSCs per individual experiment. (N = 3 mice) f, Spatial
distribution of HSCs (circles) and random dots (triangles) relative to Col.1
marking bone surfaces (left panel) and CD105+ vasculature (sinusoids, right
panel) (n = 3 mice). P values were calculated using two-tailed Kolmogorov–
Smirnov (distance distributions, left panel P = 0.1516, right panel P > 0.9999)
and one-tailed Mann–Whitney (first bin of histograms, left, HSCs: 8.56 ± 5.74,
RDs: 6.88 ± 1.94, P = 0.50; right, HSCs: 67.52 ± 10.99, random dots: 68.53 ± 3.51,
P = 0.35) tests. Data points with mean ± s.d. (red for HSCs, blue for random
dots). NS, not significant. Epi: epiphysis, meta: metaphysis, dia: diaphysis.