mRNA fold change measured by qRT-PCR for osteoblast related genes. Quantitative Real Time PCR of osteoblast related genes from rhBMP2 induced sMSC at indicated timepoints were normalized and compared according to housekeeping genes as control: mHPRT - hypoxanthine phosphoribosyltransferase 1, mGAPDH - glyceraldehyde-3-phosphate synthase and mHMBS - mouse hydroxymethylbilane synthase. A) TGFB1 relative levels were upregulated 3.2 fold after 30 min of rhBMP2 induction (p<0.001) followed by a decrease to basal levels after 2 h. B) TGFBR1 mRNA increase levels of up to 3.6 fold at 1 h and more than 4.9 fold at 2 h (Figure 6B, p<0.001). C) COL1 displays a punctual increase at 1 h after stimulus (8.6 fold, p<0.001). D) COL4A showed a progressively rising pattern at timepoints studied (5.96 fold at 1 h and 10.8 at 2 h, p<0.05 and p<0.001, respectively). E) TWIST is downregulated (0.39 fold after 10 min) but after 1h shows a slight increase (1.6 fold) at 1 h. F) SMAD2 presented a slight increase of 3.4 fold at 10 min and 2 h (p<0.001). Data represent three independent experiments and standard deviation bar is shown for all experimental timepoints studied. Asterisks represents levels of significance for statistical analysis (Two-Way ANOVA, Bonferroni test) between control and treatment groups (*p<0,05 and **p<0,01). ns: non statistically significant.
Halcsik et al. BMC Cell Biology 2013 14:47 doi:10.1186/1471-2121-14-47