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. 2014 Dec 15;105(12):1616–1625. doi: 10.1111/cas.12547

Fig. 2.

Fig. 2

Vascular endothelial growth factor (VEGF)-specific short interfering RNA (siVEGF)/CH-CA-Spe nanogel complex suppressed VEGF expression in vitro. (a) siVEGF/nanogel complex was prepared under the indicated conditions and added to Renca cell culture (0.8 μg of siRNA/10 μL/well). Twenty-four hours later, VEGF mRNA levels were evaluated by real-time RT-PCR analysis. (b) Transmission electron micrograph image of siVEGF/CH-CA-Spe nanogel complex formed at the cation/phosphate (C/P) ratio of 40 at 25°C for 30 min (concentration: 0.8 μg of siRNA/10 μL). The white small dots represent nanogel particles, and the white arrow indicates colloidal siRNA/nanogel complex with a diameter of 50–100 nm. (c and d) Renca cells were transfected with siVEGF/nanogel complex (0.8 μg of siRNA/10 μL) at different doses, while control groups were transfected with control non-silencing siRNA (siCont)/nanogel (0.8 μg of siRNA/10 μL) or left untransfected (UT). Twenty-four hours later, real-time RT-PCR (c) and ELISA (d) analyses were performed. (e) VEGF mRNA was measured by real-time RT PCR at the indicated period after transfection with siVEGF/nanogel complex (0.8 μg of siRNA/10 μL/well). (f and g) The indicated cells were transfected with siVEGF/nanogel complex (0.8 μg of siRNA/10 μL) at doses of 15 (f) and 10 (g) μL/well, and real-time RT-PCR analysis was performed 24 h later. Data represent the means ± SD (n = 3). *P < 0.05 versus siCont; **P < 0.01 versus siCont; +P < 0.001 versus siCont; ++P < 0.005 versus siCont.