STAT2 Antibody - #AF6342

Figure 5 JNJ-7706621 regulated IL-8 gene’s translation via interacting with AUF1 and inhibited tumor cells’ proliferation. (a) The remaining amount of IL8 mRNA after treated with 1.75 μM JNJ-7706621 for 24 h in SK-MEL-2 cells. Actinomycin D (5 μg/mL) was added to stop the transcription to detect remaining IL8 mRNA with qRT-PCR. The fitting curve was performed using GraphPad Prism. (b) The REMSA result of AUF1 (750 nM) incubated with the FAM-labelled RNA at 250 nM in the presence of JNJ-7706621. The sample was incubated in an incubation buffer (10 mM Tris-HCl, pH 8.0, containing 50 mM KCl, 2 mM DTT, 0.5 mM EDTA, and 0.1 µg/µL BSA) and loaded on a 6% native polyacrylamide gel. (c) The wild-type plasmid (3′WT) and mutant plasmid (3′del) were transfected in SK-MEL-2 cells. After incubation, the relative luciferase activity (RL/FL) was detected and normalized with the data of 3′del. (d) The relative luciferase activity of 3′WT and 3′WT dual-luciferase reporters after treated with JNJ-7706621 (1.75 μM), normalizing with the data of 3′WT. (e) The expression of IL8 in SK-MEL-2 cells treated with 1.75 and 3 μM JNJ-7706621. (f) The expression of IL8 in SK-MEL-2 cells treated with 80 nM siRNA of AUF1. JNJ-7706621 was added after 10-h pre-incubation with the siRNA. (g) Inhibitory effect of JNJ-7706621 in different cancer cells for 48 h in the MTT assay. All experiments were parallelly repeated in triplicate, and data are shown as means ± SEM of n = 3.