Figure 5
HuR associates with and stabilizes Atp6v0d2 mRNA. (a) The whole-cell lysates of RAW264.7 cells treated with or without poly(I:C) stimulation for 8 h were immunoprecipitated with control-IgG- or anti-HuR-antibody-conjugated beads. The precipitated mRNA was isolated and the bead-bound Atp6v0d2 mRNA was quantified with RT–qPCR. (b) Wild-type (WT) and HuR KO RAW264.7 cells were stimulated with poly(I:C) and treated with actinomycin D (2.5 µg/ml) for the indicated times. The amount of Atp6v0d2 mRNA was quantified with RT–qPCR and normalized to the level of Atp6v0d2 mRNA at time zero. (c) HEK293T cells were transiently transfected with empty or HuR-expressing plasmid and then stimulated for 8 h with poly(I:C). After the medium was changed, the cells were treated with actinomycin D (5 µg/ml) for the indicated times. The amount of Atp6v0d2 mRNA was quantified with RT–qPCR and normalized to the amount of Atp6v0d2 mRNA at time zero. Data are the means ± SE of triplicate independent experiments. *p < 0.01, **p < 0.05, Student’s t test.
