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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Nature. 2019 Jan 16;565(7740):511–515. doi: 10.1038/s41586-019-0874-3

Figure 3. Enhanced collagen hydroxylation relies on glutamine-dependent α-ketoglutarate production.

Figure 3.

(a) Intracellular α-ketoglutarate (αKG) levels in cultured chondrocytes (n=3 biologically independent samples). (b) Fractional contribution of 13C5-glutamine (Gln) to αKG (n=3 biologically independent samples). (c) Hydroxyproline (OH-Pro) content in neonatal growth plates of mice treated with BPTES, with or without αKG (n=6 for Phd2chon+-veh, Phd2chon--BPTES or Phd2chon--BPTES+αKG mice; and n=7 for Phd2chon+-BPTES, Phd2chon+-BPTES+αKG or Phd2chon--veh mice). (d) Safranin O staining of the tibia of mice treated with BPTES, with or without αKG, and quantification of the percentage Safranin O (SafO) positive matrix relative to bone volume (BV) (n=5 for Phd2chon+-BPTES mice; n=6 for Phd2chon+-veh or Phd2chon--veh mice; and n=7 for Phd2chon+-BPTES+αKG, Phd2chon--BPTES or Phd2chon--BPTES+αKG mice). Scale bar is 250 μm. Data are means ± SEM in (a, b), or means ± SD in (c,d). ***p<0.001 vs. Phd2chon+ (two-sided Student’s t-test), #p<0.05 (ANOVA), §p<0.05 vs. Phd2chon+-veh, °p<0.05 vs. Phd2chon--veh (ANOVA). Exact p values: 0.0009 (a); 0.00004 (b); Phd2chon+-veh vs. Phd2chon--veh 0.0010 (c) or 0.00001 (d); Phd2chon+-veh vs. Phd2chon+-BPTES 0.000001 (c) or 0.00003 (d); Phd2chon+-veh vs. Phd2chon--BPTES+αKG 0.005 (c) or 0.000002 (d); Phd2chon+-BPTES vs. Phd2chon+-BPTES+αKG 0.0000001 (c); Phd2chon--veh vs. Phd2chon--BPTES 0.00005 (c) or 0.0000002 (d); Phd2chon--BPTES vs. Phd2chon--BPTES+αKG 0.0003 (c).