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. 2012 Mar 23;7(3):e32989. doi: 10.1371/journal.pone.0032989

Figure 1. Hypoxia downregulates PEDF at the protein level in melanocytes and human melanoma cell lines.

Figure 1

Western blot analysis of (A) extracellular PEDF (PEDFe) protein levels in conditioned medium (CM) and (B) HIF2α protein levels in whole-cell extracts (B) from M330 primary melanocytes incubated under normoxia (21% O2) or hypoxia (1% O2) for 12 h or 24 h. β-tubulin was used as loading control. Quantitative RT-PCR analysis of (C) PEDF mRNA levels and (D) VEGF mRNA levels in M330 primary melanocytes incubated in normoxia or hypoxia for 12 h or 24 h. PEDF and VEGF mRNA levels are shown relative to cells in normoxia after normalization to β-actin. Bars represent average ± standard deviation (SD) (**P<0.01). (E) Western blot analysis of PEDFe protein levels in CM and HIF1α in whole-cell extracts from M438 primary melanocytes, SBcl2 and WM164 melanoma cell lines incubated in normoxia or hypoxia for 12 h or 24 h. β-actin was used as loading control. Quantitative RT-PCR analysis of (F) PEDF mRNA levels and (G) VEGF mRNA levels in M438 primary melanocytes, SBcl2 and WM164 melanoma cell lines incubated under normoxia or hypoxia for 12 h or 24 h. PEDF and VEGF mRNA levels are shown relative to normoxia after normalization to β-actin. Bars represent average ± SD (**P<0.01; ***P<0.001).