Figure 4

The JNK cascade is the principal pathway inhibited by Reversine and SP600125. (A–C) Gene Ontology analysis of the common target genes (22) of Reversine and SP600125 was done by the Cytoscape associated-plugin ‘ClueGO’. ‘Positive regulation of JNK cascade’ was the most associated term with both Reversine and SP600125 (A). Network view of target genes of Reversine and/or SP600125 was performed by Cytoscape and GeneMANIA associated-plugin. Red colors represent the inhibited proteins through the Kinase panel test and JNK related nodes were highlighted in green zones after Reversine (B) and SP600125 (C) treatment. (D) Overexpression and activation of JNK was investigated in RKO colon cancer cell line comparatively to the normal human colon mucosal epithelial cell line NCM460. (E) Inactivation of JNK by SP600125 or Reversine in RKO cells was confirmed by Western blot. (F) Downstream targets of JNK pathway after treatment with Reversine or SP600125 were checked by western blot.