Fig. 2. HuR regulates the expression of Snail.
A. RNP-IP detection of HuR binding RNAs of EMT related genes. Data for each individual mRNA was normalized to the IgG pull-down product of that mRNA. Bar graphs show Mean ± SEM of 9 repeats. B. Stability of Snail mRNA in MIA PaCa-2 HuR WT or HuR KO cells. Transcription was blocked by actinomycin D (5 μg/ml) treatment 30 min before the first sample was collected (0 h). Data shows Mean ± SEM of 9 repeats. C. Schematic diagram of the constructions of the full length and 2 truncations of 3’-UTR of Snail mRNA into the dual-luciferase reporter. D. Luciferase reporter assay. MIA PaCa-2 HuR KO cells were co-transfected with HuR (or vector) and the dual-luciferase reporter with Snail 3’-UTR constructions (either the Full length, AREs or ΔAREs, or empty reporter). E. Scratch assays in MIA PaCa-2 HuR-KO cells with Snail overexpression. Cells were transfected with empty vector (pVec) or Snail gene (pSnail) or 48 h before seeded at 3×105 cell/ml in 24 well plate to form monolayer. *, p<0.05; **, p<0.01 with one-way ANOVA-Tukey’s test or Student’s t-test.