Extended Data Fig. 4: Glis1 did not suppress OCR during somatic cell reprogramming.
From: Glis1 facilitates induction of pluripotency via an epigenome–metabolome–epigenome signalling cascade
a, Mitochondrial stress test using Seahorse XF24 analyzer on day 8. SKO plus Flag as a control compared to SKO plus Glis1. Flag as the control to all group. Data are presented as the mean ± S.D. (n = 3, each group). b, Oxygen consumption rates (OCR) of mitochondrial maximal respiration quantified from the mitochondrial-stress test in (a) are shown. Data are presented as the mean ± S.D. (n = 3, each group).Group differences are analyzed by the two-tailed Student’s t test. c, Mitochondrial DNA copy numbers in MEFs with SKO plus Glis1 or SKO plus Flag. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. d, Mitochondrial stress test using Seahorse XF24 analyzer on day 8. SKO plus shLuc as a control compared to SKO plus shGlis1, Flag as a control to all groups. Data are presented as the mean ± S.D. (n = 3, each group). e, Oxygen consumption rates (OCR) of mitochondrial maximal respiration quantified from the mitochondrial stress test in (d) are shown. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. f, Flow cytometry analysis of SSEA1 in MEFs transduced with SKO in reprogramming on day3. (n = 3, each group). g, SSEA1- cells were infected with retroviral Flag or Glis1. GFP+ colonies were counted on day 20. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. h, Mitochondrial stress test using Seahorse XF24 analyzer on day 8. SSEA1- plus Flag as a control compared to SSEA1- plus Glis1. Data are presented as the mean ± S.D. (n = 3, each group). i, Oxygen consumption rates (OCR) of mitochondrial maximal respiration quantified from the mitochondrial stress test in (h) are shown. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. j, Glycolysis stress test of iPSCs by Seahorse XF24 analyzer. The resultant iPSCs of SKO plus Flag as a control compared to SKO plus Glis1. Data are presented as the mean ± S.D. (n = 3, each group). k, Extracellular acidification rates (ECAR) quantified from the glycolysis stress test in (j) are shown. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. l, Mitochondrial stress test of iPSCs by Seahorse XF24 analyzer. The resultant iPSCs of SKO plus Flag as a control compared to SKO plus Glis1. Data are presented as the mean ± S.D. (n = 3, each group). m, Oxygen consumption rates (OCR) of mitochondrial maximal respiration quantified from the mitochondrial stress test in l, are shown. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. n, Relative concentration of pentose phosphate pathway (PPP) metabolites and nucleotide-related molecules. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. o, Relative band densities of H3K27Ac upon Glis1 overexpression and knockdown were quantified using Image J. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test. p, Western blot analysis of Pan Kla and H3K18la after 20 mM lactate supplementation and knockdown of Ldha. Data are presented as the mean ± S.D. n = 3 independent experiments were repeated with similar results. q, Relative band densities of Pan Kla and H3K18la upon Glis1 overexpression and knockdown were quantified using Image J. Data are presented as the mean ± S.D. (n = 3, each group). Group differences are analyzed by the two-tailed Student’s t test.
