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. 2023 Oct 4;622(7984):834–841. doi: 10.1038/s41586-023-06611-6

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© The Author(s) 2023, corrected publication 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, Expression of haemoglobin in cartilages from mice at different developmental stages by RT–qPCR. *P = 0.0104 (Hba) and 0.0179 (Hbb), and ****P < 0.0001. b,c, Expression level of the indicated genes in 4-day in vitro cultured cartilaginous tissues (b) and primary chondrocytes culture for 12 h (c). For panel b, ***P = 0.0003 (Hba), ***P = 0.0007 (Hbb), **P = 0.0029 (Hif1a), P = 0.5285 (Hif2a), **P = 0.0042 (Epo) and **P = 0.0030 (Epor). For panel c, ***P = 0.0007 (Hba), ***P = 0.0008 (Hbb), **P = 0.0016 (Hif1a), P = 0.8062 (Hif2a), **P = 0.0031 (Epo) and ***P = 0.0002 (Epor). d,e, Expression of the indicated genes in primary cultured chondrocytes upon conditional knockout (cKO) of both Hif1a and Hif2a (Hif1/2a) in either 20% (d) or 1% (e) O₂ for 6 h. For panel d, **P = 0.0017 (Hif1a), ***P = 0.0009 (Hif2a), **P = 0.0027 (Hba), **P = 0.0067 (Hbb), ***P = 0.0001 (Epo) and P = 0.1996 (Epor). For panel e, **P = 0.0065 (Hif1a), **P = 0.0018 (Hif2a), *P = 0.0236 (Hba), *P = 0.0198 (Hbb), *P = 0.0270 (Epo) and P = 0.0671 (Epor). f, Expression of the indicated genes in the primary chondrocyte culture in 20% or 1% O₂ for 6 h. **P = 0.0098 (Klf1). g,h, Expression of the indicated genes in primary cultured chondrocytes upon cKO of Klf1⁻ in either 20% (g) or 1% (h) O₂ for 6 h. For panel g, **P = 0.0030 (Klf1), **P = 0.0030 (Hba), **P = 0.0031 (Hbb), P = 0.5240 (Hif1a) and P = 0.0919 (Hif2a). For panel h, **P = 0.0014 (Klf1), **P = 0.0031 (Hba), **P = 0.0031 (Hbb), P = 0.1152 (Hif1a) and P = 0.5463 (Hif2a). i, Schematic loci of mouse globin. β_maj, β_major; β_min, β_minor; Chr., chromosome; HS, DNase I hypersensitive site; LCR, locus control region; pro., promotor. The red triangles indicate KLF1-binding sites. j,k, ChIP–qPCR results for KLF1 binding to the LCR and promoters of α-globin (j) and β-globin (k) loci in chondrocytes. P = 0.0014 (HS26), 0.0052 (Hba1-pro), 0.0027 (HS-1), 0.0087 (HS-2), 0.0078 (HS-3), 0.0023 (Hbb-bs-Pro). n = 3 biologically independent experiments (c–h,j,k) or samples (a,b). All error bars represent s.e.m. P values were calculated using one-way ANOVA test (a) or two-tailed Student’s t-test (b–h,j,k); adjustment was not made for multiple comparisons.

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