Fig. 2.

HuR increases nucleolin expression, not NCL mRNA levels. (A and B) Forty-eight hours after transfection of HeLa cells with either control (Ctrl) siRNA or HuR-directed siRNA, lysates were prepared to assess the levels of nucleolin, HuR, and loading control β-actin by Western blot analysis (A), and the levels of NCL mRNA were measured by RT-qPCR using GAPDH mRNA for normalization (B). (C and D) Twenty-four hours after transfection with a control plasmid expressing TAP or a plasmid overexpressing HuR as a fusion protein (TAP-HuR), the levels of nucleolin, HuR-TAP, endogenous (Endog.) HuR, and β-actin were assessed by Western blot analysis (C), and the levels of NCL mRNA were assessed by RT-qPCR (D), as explained in panel B. Western blotting signals in panels A and C were quantified by densitometry, and the relative abundance of nucleolin was calculated. Data in panels A to D represent the means of 3 independent experiments. s.d., standard deviation.