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. 2022 Nov 11;5:1230. doi: 10.1038/s42003-022-04156-4

Fig. 7. The pentose phosphate pathway is required for proper hypertrophy and bone growth in ex vivo metatarsal culture.

Fig. 7

a WT metatarsal explants cultured for 4 days in the presence or absence of 6-AN. Representative of n = 6–11 bones per group. b Quantification of metatarsal and distal cartilage growth from the beginning of the culture period (as percent of initial length of metatarsal or distal cartilage). n = 6–11 bones per group. c Toluidine blue staining for morphology and IHC for collagen X (hypertrophic marker) in ex vivo-cultured WT metatarsals treated with 6-AN. Representative of n = 3–5 bones per group. CZ columnar zone. HZ hypertrophic zone. d Measurements of the lengths of the CZ and HZ, and the ratio of the CZ to HZ, in ex vivo-cultured metatarsals treated with 6-AN. e Igf2 null metatarsal explants cultured for 4 days with and without 6-AN. Representative of n = 6–12 bones per group. f Quantification of metatarsal and distal cartilage growth from the beginning of the culture period (as percent of initial length of metatarsal or distal cartilage). n = 6–12 bones per group. g Toluidine blue staining for morphology and IHC for collagen X in Igf2 null metatarsals treated with 6-AN. Representative of n = 3–6 bones per group. h Measurements of the lengths of the CZ and HZ, and the ratio of the CZ to HZ, in Igf2 null metatarsals treated with 6-AN. n = 3–6 bones per group. Scale bar: 100 µm. Data are presented as individual measurements with mean + SEM. Note that the control data (0 µg/mL 6-AN) for b, d, f, and h are also shown in Fig. 5. Statistical significance was calculated by Brown–Forsythe and Welch ANOVAs with Dunnett’s T3 test for multiple comparisons (b) or one-way ANOVA with Tukey’s test for multiple comparisons (d, f, h). p-values indicated as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.