Fig. 5.

miR-200a promoted JNK2 protein translation in human BC cells. a Total RNA was extracted from the cells and JNK2 mRNA levels were evaluated using real-time PCR. b Indicated cells were treated with cycloheximide (CHX) for denoted times and then cell extracts were subjected to western blot for determination of JNK2 protein degradation. β-Actin was used as protein loading control. c After treatment with MG132 (10 μM) for 30 min, newly-synthesized JNK2 protein in T24T(Vector) and T24T(miR-200a) cells was monitored by pulse assay using [³⁵S]-labeled methionine/cysteine. WCL indicates whole cell lysate. Coomassie blue staining was used for protein loading control. d Indicated stable transfectants, T24T(Vector) and T24T(miR-200a) were used to analyze the indicated protein levels using western blot. Results shown are representative of three independent experiments. e shS6 and Nonsense control were stably transfected into T24T(miR-200a) cells and transfectants were identified by western blot. β-Actin was used as protein loading control. Results shown are representative of three independent experiments