Fig. 6

NUAK1 activates the JNK/c-Jun pathway to derive Slug-mediated migration and invasion in ESCC cells. A–C Western blot analysis of p-c-Jun (S73) and c-Jun (A), p-Akt (S473) and Akt (B), p-JNK (T183/Y185), JNK, p-Erk (T202/Y204), Erk, p-p38 (T180/Y182) and p38 (C) in NUAK1-overexpressing EC109 and KYSE510 cells, NUAK1-knockdown KYSE30 and KYSE150 cells and control cells. D KYSE30 cell lysates were subjected to immunoprecipitation with IgG, anti-NUAK1 (left panel) or anti-JNK (right panel) antibodies. Immunoprecipitates were blotted with indicated antibodies. E The expression of p-c-Jun (S73), c-Jun and Slug was analyzed in NUAK1-overexpressing EC109 cells treated with SP600125 and JNK-IN-8. Quantification of indicated protein levels of n = 3 independent biological experiments (right panel). G The promoter activity of Slug was analyzed using the luciferase assay in NUAK1-overexpressing EC109 cells treated with SP600125 and JNK-IN-8. H, Cell migration and invasion were analyzed in NUAK1-overexpressing EC109 cells treated with SP600125 and JNK-IN-8. Error bars denote mean ± SD. **P < 0.01; ***P < 0.001. P value was calculated by one-way ANOVA with post hoc intergroup comparison by the Tukey’s test (for F–H)