Fig 2.

Regulation of PEDF in human cardiac cells. HACM (A) and HACF (B) were cultivated under normoxic (control) and anoxic conditions or stimulated with CoCl₂ (0.1–100 μM), DP (0.1–100 μM) or DMOG (1 mM to 1 μM) for 48 hrs; PEDF protein in conditioned media was determined by specific ELISA. Values are given as ng/100,000 cells and represent mean values ± S.D. of three independent determinations (A, B). mRNA was isolated from HACM and HACF cultivated for 8 hrs under normoxic (control) or anoxic conditions or stimulated with 100 μM CoCl₂, 100 μM DP or 1 mM DMOG. Real-time PCR for PEDF was performed employing specific primers. Values, given as x-fold of control, represent mean values ± S.D. of three independent determinations and were normalized to GAPDH levels (C). HACM and HACF were cultivated for 6 hrs under normoxic (control) or anoxic conditions or stimulated with 100 μM CoCl₂, 100 μM DP or 1 mM DMOG; nuclear extract was isolated and HIF-1α activation was measured using a specific transcription factor ELISA. Values represent mean values ± S.D. of three independent determinations and are given as x-fold of control (D). *P < 0.005, ^#P < 0.05.