Fig. 6

MEIS2C/D interacted with β-catenin and YAP in different modes. a The rescue MEIS2C or MEIS2D cDNA clone, which is resistant to siRNA interference, were used to rescue the MEIS2C/D expression after MEIS2C/D knockdown in HCC-LM3 cells. The level of phospho-LATS1, phospho-YAP, phospho-CDC73 and total LATS1, YAP, CDC73 and SHP2 were detected in MEIS2C and MEIS2D rescued HCC-LM3 cells, respectively. b Immunofluorescence staining for DAPI (Blue), YAP (Green) in HCC-LM3 cells. c Lysates were sequentially immune-precipitated (IP) from HCC-LM3 cells transfected with Flag-MEIS2C (left) or Flag-MEIS2D (right) and immunoblotted by CDC73, β-catenin and YAP antibody. d Immunofluorescence images showing dual staining for CDC73 (Red) with Flag-MEIS2C or Flag-MEIS2D (Green) in HCC-LM3C cells. Merged picture with sites of co-localization in yellow are shown. e SHP2 constructs [wildtype (WT), dominant negative (DN, C463S mutant), and constitutively active (CA, E76K mutant)] and CDC73-Myc co-transfected with Flag-MEIS2C or Flag-MEIS2D in HEK293T cells. MEIS2 were immune-precipitated from whole cell lysates and probed with Myc antibody to check the interaction with CDC73. f Phosphorylation-resistant (PR) CDC73-PR-Myc mutant (CDC73-Y290/293F/315F-mutant) vector was transfected together with either Flag-MEIS2C or Flag-MEIS2D vector into HEK293T cells. Anti-Myc immune-precipitates were analyzed by immunoblotting with anti-Myc and anti-Flag antibodies