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

Figure 1

The p5 internalization and release by hADMSCs. (a) The internalization of p5 was analyzed by fluorescence microscopy in hADMSCs primed 1, 2, 4, 8, or 24 hours with 3 μg/mL p5 (green). Cells were also observed after 24 hours priming on Days 2, 3, 4, 5, 10, and 14. (b) The internalization of p5 was analyzed by fluorescence microscopy in BAECs primed with different doses of p5 (green) for 24 hours. The intensity of p5 staining in cytoplasm was correlated with the dosages of p5. (c) Subconfluent cultures (2 × 10⁵) of adherent hADMSCs were exposed to 12 μg/mL biotinylated p5 for 24 hours. After several washes and trypsinization, p5-primed hADMSCs were further cultured and their conditioned medium (CM) was collected every 24 hours. The internalization of p5 was analyzed by fluorescence microscopy in BAECs treated with p5-primed hADMSCs CM for 24 hours. Scale bar: 100 μm. (d) The fluorescein staining intensities of p5 in hADMSCs in (a) were quantified in the bar graph as means ± SD (error bars). Significance (denoted as ) was calculated compared to the untreated control group using Student’s -test (with , ). (e) The fluorescein staining intensities of p5 in BAECs in (b) were quantified in the bar graph as means ± SD (error bars). (f) The fluorescein staining intensities of p5 in BAECs in (c) were quantified in the bar graph as means ± SD (error bars). Significance (denoted as ) was calculated compared to the untreated control group using Student’s -test (with , ).

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