Figure 1. Depletion of km23-1 blocks constitutive ERK activation in human CRC cells.

A. EV, NC siRNA, and km23-1 siRNA stable transfectants (clones 1 and 5) were used to isolate RNA. RT-PCR was performed and products were analyzed as described in the “Materials and Methods.” The data plotted are the mean ± SE of three independent experiments. *p<0.01 compared to the NC siRNA. B: Western blotting of phospho- and total protein expression levels for ERK1/2 in RKO human CRC cells. Bottom, DIC protein was assessed as a loading control. Data are representative of three independent experiments. C: Western blotting of phospho- and total protein expression levels for ERK1/2 in HCT116 cells stably transduced with the lentiviral particles described in the “Materials and Methods.” Top, confirms knockdown of endogenous km23-1. Bottom, GAPDH protein was assessed as a loading control. Data are representative of three independent experiments. D: CBS cells were infected with either pilenti-NC siRNA-GFP or pilenti-km23-1 siRNA-GFP pools. 24 h after infection, Western blotting was performed using the indicated antibodies. Top, confirms knockdown of endogenous km23-1. Bottom, DIC protein was assessed as a loading control. Three independent experiments were performed and representative figures are shown.