Effect of higher doses of cytokines on KGFR mRNA and protein expression in MCF-7 cells.

<p>(<b>A–B</b>) Quantitative real-time PCR analysis of KGFR mRNA expression in MCF-7 cells following treatment with 100 ng/ml IL1β, IL2 or TNFα for 24 and 48 h. Relative KGFR mRNA levels are shown as fold value of the level of KGFR mRNA in untreated cells. Each experiment was performed in triplicate, and mRNA levels were normalized to GAPDH mRNA expression. Error bars represent standard deviations. *<i>P</i><0.01. (<b>C–D</b>) Western blot analysis of KGFR protein levels in MCF-7 cells untreated or treated with 100 ng/ml IL1β, IL2 and TNFα for 24 and 48 h. KGFR protein expression was evaluated by blotting with an anti-Bek antibody. Western blot with anti-Tubulin antibody was used as loading control. The images are representative of at least three independent experiments. The intensity of the bands was evaluated by densitometric analysis, normalized and reported as relative expression with respect to the untreated cells. Densitometric analysis was also performed for each experiment and reported as a graph. Error bars represent standard deviations. #<i>P</i><0.05.