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. 2019 Jan 8;7:2. doi: 10.1038/s41413-018-0038-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — High-throughput phenotypic screening (HTS) identifies high cortical bone thickness in Notum^−/− mice. a, b Histograms showing distribution of midshaft femur cortical bone thickness of N = 13 845 mice from 3 366 distinct global gene knockout mouse lines and trabecular bone volume fraction (BV/TV) in the fifth lumbar vertebral body of N = 13 979 mice from 3 399 distinct global gene knockout mouse lines. Mean values are normally distributed. HTS data for selected genes with substantially increased or decreased values are indicated by arrows. c–g 16-week-old Notum⁻/⁻, Notum^+/− and WT mice. c–f Midshaft femur cortical total bone area, bone area, cortical thickness, polar MOI (N males: WT = 23, Notum^+/−= 50, Notum^−/−= 17; females: WT = 22, Notum^+/−= 57, Notum^−/−= 18). g Femur shaft bone strength (maximum load; Max. load) measured by 4-point bending (N males: WT = 23, Notum^+/−= 20, Notum^−/−= 17; N females: WT = 21, Notum^+/−= 20, Notum^−/−= 18). h Midshaft femur cortical bone thickness in male and female Notum^−/− and WT mice at different ages (N males: WT: 4 weeks = 14, 16 weeks = 10, 32 weeks = 6, 52 weeks = 6; N Notum^−/−: 4 weeks = 13, 16 weeks = 10, 32 weeks = 6, 52 weeks = 8; N females: WT: 4 weeks = 9, 16 weeks = 8, 26 weeks = 13, 43 weeks = 13; N Notum^−/−: 4 weeks = 10, 16 weeks = 9, 26 weeks = 14, 43 weeks = 9). i Vertebral trabecular BV/TV (central part of vertebral body; LV5) of 16-week-old male mice (N: WT = 23, Notum^−/− = 17). j Effect of ovariectomy (OVX) on midshaft femur cortical bone thickness in WT and Notum^−/− mice. OVX or sham-surgery was performed at 18 weeks of age and bones were analyzed 8 weeks later (N WT: sham = 13, OVX = 13; N Notum^−/−: sham = 14, OVX = 16), lean ^P < 0.01 OVX vs sham. k Notum mRNA levels in different tissues in female mice. Arbitrary levels are given with liver indicated as 100%. **P < 0.01 vs tibia diaphyseal bone without marrow (N = 10–11). i Notum mRNA in primary cultured mouse osteoblasts (Obl), pre-osteoclasts (preOcl) and osteoclasts (Ocl). BMP-2 is indicated for BMP-2 stimulated osteoblasts. Arbitrary levels are given with unstimulated Obl indicated as 100% with four cell cultures per data point