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Figure 5


Fig. 5. hCAF1 regulates asymmetric dimethylation of proteins in MCF-7 cells. (A) Lysates of MCF-7 cells transfected either with control (scramble) or specific hCAF1 or PRMT1 siRNA duplexes were analyzed by western blotting using the pan antibody ASYM24. Results are representative of several individual experiments. (B) Quantification of arginine dimethylated bands revealed with ASYM24 antibody. Data represent the changes in arginine dimethylated bands after hCAF1 and PRMT1 knockdown, expressed as the ratio of the quantity present in knockdown cells to that found in cells treated with scrambled control siRNA. Data representing the mean of three independent experiments are shown with error bars. (C) Western blot analysis showing the expression of the indicated proteins in MCF-7 cell extracts after transfection with control (scramble) or hCAF1 or PRMT1 siRNA duplexes. GAPDH served as a loading control. (D) Confocal fluorescence microscopy experiments showing the subcellular distribution of arginine dimethylated proteins in MCF-7 cells transfected either with control (scramble) (a) or specific hCAF1 (b) or PRMT1 (c) siRNA duplexes revealed with ASYM24 antibody.





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