Figure 3.

Importance of the uORF for A3G mRNA expression. (A) Schematic representation of the different A3G 5′-UTR constructs used in this study. (B) HEK 293T cells were transfected with wt or mutated A3G constructs. Proteins were separated by SDS/PAGE and analyzed by immunoblotting with anti-A3G (#9968) and anti-β-actin (#A5316) antibodies. Bands were quantified using ImageJ, normalized to actin, and relative expression of A3G is represented in a histogram (orange bars). Total RNA was extracted from transfected cells and RTqPCR was performed to study the relative expression of wt and mutated A3G constructs (blue bars). The ratio protein/RNA is indicated below. Data represent the mean ± S.E.M. for at least three independent experiments. p-values are indicated as follows: * <0.05, ** <0.01, *** <0.001.