Figure 6: BMP-2-mediated regulation of myosin Va may be modulated by Erk signaling with Ca2+ involvement. (a) IPA software analysis of the relationships and interactions between BMP-2, Erk1/2, myosin Va, and actin. Although communication between the genes of these proteins genes has previously been reported, heretofore, a direct interaction between BMP-2 and myosin Va was not clarified. (b) Western demonstrates the presence of key proteins involved with myosin Va and Erk signaling in the lysates of rat VSMCs either adenovirally transfected to overexpress BMP-2 or treated with exogenous BMP-2 (250 ng/mL) for 48 hours. (c) qRT-PCR analysis demonstrating myosin Va gene mRNA was upregulated in VSMCs treated with 250 ng/mL BMP2 for 48 hours and downregulated in the presence of Erk1/2 inhibitor U0126 (dose 5 μM). (d) Migration of VSMCs subjected to identical conditions was detected by Boyden chamber assay (BMP-2+ alone: versus BMP-2+/U0126+: ). (e) Oscillatory increase of intracellular Ca2+ concentration ( ) within individual cells of the colony. Chronological image sequence goes from left to right at 1.5-second intervals over 5 minutes. (f) Oscillations exhibited a mean interval of intracellular Ca2+ concentration (BMP-2 concentration 250 ng/mL, ). (g) Fluorescence ( ) of individual cells, values represent mean ± SD; *Student’s -test: ).