Figure 2

Loss of SMYD3 diminishes hypoxia signaling. A, immunoblotting of indicated proteins in SMYD3-deficient or wildtype HEK293T cells (SMYD3^−/− or SMYD3^+/+). B and C, qPCR analysis of GLUT1 (B) and PGK1 (C) mRNA in SMYD3-deficient or wildtype HEK293T cells (SMYD3^−/− or SMYD3^+/+) under normoxia (21% O₂) or hypoxia (1% O₂) for 24 h. Data show mean ± SD; Student’s two-tailed t test. ∗p < 0.05, ∗∗p < 0.01. Data from three independent experiments. D–F, qPCR analysis of GLUT1 (D), PDK1 (E), and BNIP3 (F) mRNA in SMYD3-deficient or wildtype HEK293T cells (SMYD3^−/− or SMYD3^+/+) treated with or without CoCl₂ (200 μM) for 8 h. Data show mean ± SD; Student’s two-tailed t test. ∗∗p < 0.01, ∗∗∗∗p < 0.0001. Data from three independent experiments. G, immunoblotting of indicated proteins in Smyd3-deficient or wildtype MEF cells (Smyd3^−/− or Smyd3^+/+). H and I, qPCR analysis of Glut1 (H) and Pgk1 (I) mRNA in Smyd3-deficient or wildtype MEF cells (Smyd3^−/− or Smyd3^+/+) under normoxia (21% O₂) or hypoxia (1% O₂) for 24 h. Data show mean ± SD; Student’s two-tailed t test. ∗∗∗p < 0.001. Data from three independent experiments. J, immunoblotting of indicated proteins in Smyd3-deficient MEF cells reconstituted with or without wildtype Smyd3 by lentivirus. K and L, qPCR analysis of Pgk1 (K) and Vegf (L) mRNA in Smyd3-deficient MEF cells reconstituted with or without wildtype Smyd3 by lentivirus under normoxia (21% O₂) or hypoxia (1% O₂) for 24 h. Data show mean ± SD; Student’s two-tailed t test. ∗p < 0.05. Data from three independent experiments. qPCR, quantitative RT–PCR; MEF, mouse embryonic fibroblast.