Figure 2

ING4 expression in osteosarcoma cells.

Osteosarcoma cells U-2OS were transfected with pEGFP-ING4 expression vector and pEGFP-C2 control vector, respectively. Stable clones were selected with kanamycin and checked under fluorescence microscopy (A: Un-transfected U-2OS cells; B: U-2OS cells transfected with pEGFP-C2 control vector; C: U-2OS cells transfected with pEGFP-ING4 vector). D: The mRNA expression level in U-2OS cells transfected with ING4 was increased significantly in comparison to the control vector group and un-transfected group quantified by qRT-PCR method. E: The ING4 protein expression level in U-2OS cells transfected with ING4 was increased significantly in comparison to the control vector group and un-transfected group analyzed by Western blot method. ING4 mRNA expression in the stably ING4-transfected U-2OS cells was increased significantly in comparison to the control vector group and un-transfected cells (D, p < 0.01). Blots for different proteins were cropped and vertically stacked into one image with white area separated in between different proteins. Blue arrows indicate the horizontal cropping lines. All gels were run under the same experimental conditions. All data represent the mean ± standard deviation of three independent experiments. **: compared to the un-transfected group (p < 0.01), ##: compared to the cells transfected with control vector (p < 0.01).