Fig. 5.

SERCA2 and miR-29c expression in physiological hypoxia. A–D: analysis of HL-1 cells exposed to 1% O₂ for up to 24 h compared with normal oxygen conditions (N). A: Western blot of hypoxia-inducible factor (HIF)-1α expression. β-Actin was used as a loading control. B: real-time PCR analysis of Ca²⁺ handling and HIF-1 target genes. Fold changes are shown compared with normoxia. With the use of one-way ANOVA with Tukey’s multiple comparisons, the overall P values for Egln1, carbonic anhydrase 9 (Car9), SERCA2, ryanodine receptor 2 (RyR2), and phospholamban (Pln) were P = 0.0011, P = 0.0004, P = 0.0014, P = 0.033, and P = 0.0076, respectively. *P < 0.05, **P < 0.01, and ***P < 0.001 for between-group comparisons. C: Western blot of SERCA2 expression. β-Actin was used as a loading control and for normalization. Right: quantification of SERCA2 protein with normal oxygen (N) set as 100%. D: real-time PCR analysis of miR-29c expression. For A–D, all experiments were performed in triplicate. For B and D, fold changes are shown compared with normal oxygen conditions. E: miR-29c expression in mouse hearts 3 days after myocardial infarction (MI). Ischemic free wall (I) and remote (R) fractions of infarcted hearts were compared with similar tissue from sham-operated mice. n = 8 sham and n = 6 MI. Fold changes are shown compared with sham I values. For C–E, the overall P values using one-way ANOVA were P = 0.08, P = 0.08, and P = 0.0078. For C and E, *P < 0.05 by a Student’s t-test.