Effect of JNK-1 inhibition in miR-21 and PDCD4 expression.

<p>A2780CP20 cells were treated with 10 µM SP600125. (<b>A</b>) Western blot shown the inhibition of p-c-Jun after treatment with SP600125 in A2780CP20 cells compared to control (DMSO). (<b>B</b>) SYBR-I-based real-time PCR was performed to calculate the relative pre-mir-21 expression in A2780CP20 cells after treatment with SP600125 inhibitor. (<b>C</b>) Western blot and densitometric analysis of PDCD4 protein expression levels after treatment of A2780CP20 cells with SP600125. (<b>D</b>) PDCD4 protein expression levels after transfection of A2780CP20 with miR-21 oligonucleotide inhibitor. (<b>E</b>) A2780 CP20 cells were transiently transfected with two c-Jun-targeted siRNAs as described in the “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097094#s2" target="_blank">Materials and Methods</a>” section. Western blot analysis shows that both c-Jun-siRNAs decreased the c-Jun levels. SYBR-I-based real-time PCR was performed (see “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0097094#s2" target="_blank">Materials and Methods</a>” section) to calculate the relative pre-mir-21 expression levels in A2780CP20 cells after siRNA-mediated c-Jun silencing. *p<0.05, **p<0.01, ***p<0.001 compared to control. Columns represent the means of triplicates ± S.E.M.