Mesenchymal Stem Cells Loaded with p5, Derived from CDK5 Activator p35, Inhibit Calcium-Induced CDK5 Activation in Endothelial Cells

Figure 3

The activation of CDK5 by calpain in cultured BAECs. (a) The cultured BAECs were treated with 5 μM calcium ionophore A23187 and 2.5 mM CaCl₂ for 0, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or 4 hours. At the end of the treatment, the cells were collected, suspended in SDS-PAGE sample buffer, and boiled for 5 minutes. Each sample was run on a 10% SDS-PAGE gel and blotted with anti-p35, anti-phospho-CDK5 (pTyr¹⁵), anti-phospho-ERK1/2, anti-phospho-p53 (pSer¹⁵), anti-active caspase-3, anti-CDK5, anti-ERK1/2, anti-p53, anti-procaspase-3, and anti-tubulin antibodies. The first lane was loaded with the extract from untreated BAECs as control. The cultured BAECs treated with 10 ng/mL FGF2 for 10 minutes were used as positive control for the phosphorylation of ERK1/2 (lane 8). Blots shown are one of at least three independent experiments performed. (b) Densitometric analysis of immunoblots quantification. Data are means ± SD (error bars). Significance (denoted as ) was calculated compared to the untreated control (lane 1) using Student’s -test (with , ).

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