Figure 4
Effect of C/EBPβ knockdown on VEFG mRNA expression. (A) KGN cells were treated with or without cAMP (0.5 mM) for 24 h. The relative mRNA levels of VEGF were assessed by qPCR. GAPDH was used as an internal control. Data were shown as a ratio of the control treatment sample. Each value represents the mean ± SEM of 3 independent incubations. a, P < 0.05 vs. control sample. mRNA expression was also assessed by RT-PCR. The images of representative gels are shown. (B) KGN cells were transfected with a siRNA targeted against C/EBPβ or with a nontargeting siRNA as a control. After 24 h of transfection, cells were treated with or without cAMP for 24 h. Whole cell lysates were prepared and protein levels of C/EBPβ were examined by Western blotting to confirm the C/EBPβ knockdown. Protein levels of β−tubulin were also assessed to ensure equal amount of proteins. Western blotting was repeated 3 times, and the representative immunoblots are shown. Quantification of bands were performed by using ImageJ and normalized with β−tubulin levels. Data were shown as a ratio of the control treatment sample. Each value represents the mean ± SEM of 3 independent experiments. a, P < 0.05 vs. control treatment; b, P < 0.05 vs. cAMP treatment in the control siRNA. (C) The relative mRNA levels of VEGF were assessed by qPCR. GAPDH was used as an internal control. Data were shown as a ratio of the control treatment in the control siRNA. Each value represents the mean ± SEM of 3 independent incubations. a, P < 0.05 vs. control treatment in the control siRNA. b, P < 0.05 vs. cAMP treatment in the control siRNA. mRNA expression was also analyzed by RT-PCR. All PCR products were electrophoresed on the same gel and the ethidium bromide-stained gel is a representative of 3 independent incubations.
