FIGURE 2.

Vasoactive intestinal peptide (VIP) upregulates the expression of β‐catenin and Runx2. CTNNB1 (β‐catenin) (A) and RUNX2 (Runx2) (B) mRNA expression levels after 1, 4, or 7 days of bone marrow‐human mesenchymal stem cells (BM‐hMSCs) osteogenic induction in absence or presence of VIP 10⁻⁸ M was determined by real‐time PCR. Results are expressed as relative mRNA expression (relative to GAPDH levels) and referenced to undifferentiated cells (BM‐hMSCs). The means ± SEM of triplicate determinations of four independent experiments are shown. β‐catenin and Runx2 protein quantification at Day 7 of differentiation was determined by Western blot (C). Picture is a representative example of six experiments with similar results. (D) Relative expression levels of protein bands were calculated with the densitometry values of the band corresponding to the protein of interest relative to GAPDH and referenced to basal levels. Mann–Whitney U was performed (*p < 0.05; **p < 0.01; ***p < 0.001).