Figure 2.

miR-195 Improved Endothelial Functions

(A) Protein markers in miR-195-transfected and oxLDL-treated HUVECs. We used oxLDL to decrease endothelial functions, and therefore the beneficial effects of miR-195 could be detected. Quantitative data from the western blot are shown at right. See also Figures S4 and S5. (B) HUVECs were co-cultured with microRNA-transfected (50 nmol/L) and oxLDL-treated HASMCs for 5 h, and the adhesion test was performed using THP-1 cells. Left: results obtained using a fluorescence microscope with 100× magnification; right: quantitative data. Scale bar, 100 μm. Lipofectamine 2000 was used as a transfection reagent. (C) HUVECs were transfected with CD40-shRNA (1 μg/mL), and THP-1 cells were added to the HUVEC culture for the adhesion assay at 5 h post-transfection. Images were obtained using a Leica DMI6000B microscope with 100× magnification. Scale bar, 100 μm. (D) Western blot for adhesion molecules, eNOS, and the NF-κB-related molecules was measured at 48 h post-transfection of shRNA. (E) Rat neural stem cells (NSCs) isolated from the subventricular zone (SVZ) were first confirmed by the positive staining of Nestin (red in cytoplasm) and Sox2 (green in nucleus and cytoplasm). Transfection of miR-195 by HiPerFect transfection reagent increased the proliferation of NSC at 72 h. Images were obtained using a Leica DMI6000B microscope with 400× magnification. Scale bar, 20 μm. Values are presented as mean ± SEM from three independent experiments performed in triplicate. *p < 0.05, **p < 0.01, and ***p < 0.001. NC-miR, normal control microRNA. All values were expressed as mean ± SE.