Extended Data Figure 8. Vhl^iΔEC mutants show increased bone mass.

a-b, Serum alkaline phosphate levels in Hif1a^iΔEC (a) and Vhl^iΔEC (b) mutants. Data represent mean±s.e.m (n=5-6 mice for Hif1a^iΔEC mice from three independent experiments; n=6 mice for Vhl^iΔEC from three independent experiments). P values, twotailed unpaired t-test.

c, Representative μ-CT images of tibias from Vhl^iΔEC mutants and littermate controls.

d-g, Quantitative μ-CT analysis of relative bone volume (d), trabecular thickness (e) trabecular number (f), and trabecular separation (g) in proximal tibia from Vhl^iΔEC mutants and their littermate controls. Data represent mean±s.e.m (n=4 mice from from two independent experiments). P values, two-tailed unpaired t-test. Note increased bone mass in Vhl^iΔEC mutants.

h, Calcein double labelling of 5 week-old Vhl^iΔEC mutant and littermate control tibiae.

i-j, Quantitative analysis of bone formation parameters. Mineral apposition rate [MAR; (i)] and bone formation rate/bone surface [BFR/BS; (j)] for Vhl^iΔEC mutants and controls. Data represent mean±s.e.m (n=6-7 mice from three independent experiments). P values, two-tailed unpaired t-test.

k, Representative confocal images showing Calcitonin receptor staining (osteoclasts) in tibia sections from Vhl^iΔEC mutants and littermate controls. Nuclei, DAPI (blue).

l-m, Histological analysis of Vhl^iΔEC and control tibiae showing osteoclast number/bone perimeter [No. Oc./B. Pm; (l)] and osteoclast surface/bone surface [Oc. S/B. S; (m)]. Data represent mean±s.e.m (n=6 mice from three independent experiments). P values, two-tailed unpaired t-test.