Fig. 7

PKN2 suppressesErk1/2 by directly binding to and activating DUSP6. a Protein expression of DUSP6 in human colon cancer tissue array was detected by immunohistochemistry staining. The correlation between DUSP6 expression score and PKN2 expression score was explored. b Comparison of the progression free survival (PFS) of patients with different levels of PKN2/DUSP6 expression using the Kaplan-Meier method. c RKO cells were transfected with PKN2-WT-HA, siCTL, siDUSP6-1 and siDUSP6-2, separately.Western blotting was used to detect the indicated proteins. d The DUSP6 activity was detected in stable clones of SW480, HCT116 and HT-29 cells as indicated in Fig. 3. **, P < 0.01; ***, P < 0.001 versus shCTL. *, P < 0.05 versus Vector. e The 293 T cells were transfected with PKN2-WT-HA, PKN2-K686R-HA, DUSP6-WT-flag or control vector as indicated and treated with/without cardiolipin (50 μg/mL). Whole lysates pulled down with anti-flag or anti-HA antibodies were assayed for DUSP6 and PKN2 activity, respectively. For PKN2 activity assay, *, P < 0.05 versus lane 5; #, P < 0.05 versus lane 6. For DUSP6 activity assay, ***, P < 0.001 versus lane 3; ###, P < 0.001 versus lane 4. f Co-IP of DUSP6 and PKN2 in HCT116, SW480 and HT-29 cells. g Co-expression of flag-tagged DUSP6 with or without different amounts (0, 3 and 6 μg) of HA-tagged PKN2-WT in 293 T cells. h, i Flag-tagged full-length DUSP6 and its truncation mutant vector were co-expressed with HA-tagged PKN2 in 293 T cells and immunoprecipitated by anti-flag antibody. j Schematic overview on the mechanisms by which PKN2 modulates TAMs polarization and inhibits colon cancer growth