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Figure 4 | Molecular Cancer

Figure 4

From: Analysis of an alternative human CD133 promoter reveals the implication of Ras/ERK pathway in tumor stem-like hallmarks

Figure 4

Inhibition of MEK/ERK pathway attenuates CD133 expression and side population frequency. A, Inhibition of endogenous ERK activation by U0126. Immunoblot analysis was performed with phospho-ERK1/2 and ERK1/2 antibodies. B, Effects of U0126 on P5 promoter activity. Cells treated with DMSO or U0126 for 24 hours were transfected with P5 reporter plasmid, and incubated for an additional 48 hours in the presence of U0126. Data are means ± s.d. of values from three independent experiments. *P < 0.05. C, Effect of U0126 on CD133 protein expression. Caco-2 and Fuji cells were incubated with DMSO or U0126 for 72 hours, and the expression of CD133 protein was analyzed. Actin is an internal control. D, Effect of U0126 on the side population frequency in Caco-2 cells. Caco-2 cells were incubated with DMSO or U0126 for 72 hours, stained with Hoechst 33342, and subjected to side population analysis. Hoechst dye was excited with the UV laser and its fluorescence was resolved with a Hoechst blue (y-axis) and Hoechst red (x-axis). Red boxes are defined as SP fractions with low intensity of Hoechst staining. F. Bar graph representation of the percentages of side population cells in Caco-2 cells. Data are expressed as the means ± s.d. for three independent experiments. *P < 0.01.

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