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. 2016 Feb 16;113(9):E1226–E1235. doi: 10.1073/pnas.1600813113

Fig. S4.

Fig. S4.

MEKK2 regulates β-catenin stability via S675 phosphorylation. (A) C3H10T1/2 cells were transfected with different amounts of β-catenin (WT or S675A mutant) along with TOPflash-luciferase and Renilla. Luciferase activity was measured 48 h after transfection and normalized to Renilla. Expression of β-catenin was analyzed by immunoblotting with anti–β-catenin antibody. (B) β-Catenin–deficient COBs were reconstituted with Flag–β-catenin WT or S675A mutant via lentivirus-mediated delivery and were cultured under osteoblast differentiation conditions for 7 d. Transcript levels of Flag–β-catenin were measured by RT-PCR. P values for Bonferroni-corrected two-way Student’s t tests are indicated: **P < 0.01; N.S., not significant. (C) HEK293 cells were transfected with Flag–β-catenin (WT or S675A mutant) along with HA-MEKK2, and β-catenin stability was determined by pulse-chase labeling with [35S]methionine followed by autoradiography and analysis in ImageJ.