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. 2013 Nov 4;8(11):e80315. doi: 10.1371/journal.pone.0080315

Figure 4. The expression of genes implicated in osteoclasts modulation is not altered in PC3 cells by co-culturing with osteoblasts.

Figure 4

PC3 cells were pre-treated with hemin (80 μM, 24 h, black columns) or not (control, white columns) and co-cultured or not with PMOs. Total RNA was extracted and PTHrP (A), uPA (B) and TGF-β1 (C) mRNA levels were analyzed by RT-qPCR. Data were normalized to β-actin and were expressed as fold induction respect to PC3. Gelatin zymography was done to assess MMP9 activity on conditioned media from PC3 grown alone (PC3), pre-treated with hemin (PC3 Hem) and from PMOs grown alone (PMO) and co-cultured with PC3 pre-treated or not with hemin (PMO PC3 Hem and PMO PC3, respectively). Clear bands were quantified using ImageJ 1.37.v software (NIH) and normalized to total protein. One representative from at least three independent experiments is shown (NS: No significant difference; R.V.: relative values).