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. 2016 Dec 6;7:13601. doi: 10.1038/ncomms13601

Figure 6. Alendronate stimulates endothelial Notch signalling and flow in bone.

Figure 6

(a) Graph showing age-dependent changes in EdU+ ECs and bud structures in distal tibia. Blood flow measurements in the same age groups indicate a similar pattern. Data represent mean±s.d. (n=5 biological replicates). P values, two-tailed unpaired t test. (b) Tile scan images of tibia from control and Alendronate-treated 80-week-old mice. Sections were immunostained for CD31 (green) and Emcn (red). Note appearance of CD31+ blood vessels (arrows) in Alendronate-treated metaphysis. (c) Vascular front in sections of control and Alendronate-treated aged tibia immunostained for CD31 (green), Emcn (red) and Osterix (Osx, white). Nuclei, DAPI (blue). Inset shows CD31+ blood vessels in close proximity to osteoblasts. (d) Confocal images showing metaphysis of aged mice after Alendronate administration. Immunostaining shows endothelial Dll4 (green). Nuclei, DAPI (blue). (e) qPCR analysis of Dll4, Hes1, Hes5, Hey1 and Jag1 transcripts expression in ECs sorted from control and Alendronate-treated 80-week-old long bone. Data represent mean±s.d. (n=6 biological replicates). P values, two-tailed unpaired t test. (f) Increased blood flow in tibia of Alendronate-treated aged (80-week-old) mice. Data represent mean±s.d. (n=6 biological replicates). P values, two-tailed unpaired t test. (g) Blood flow measurements in tibia after short-term treatment with Alendronate. Data represent mean±s.d. (n=6 biological replicates). P values, two-tailed unpaired t test.