FIG 2.

WWOX interacts with and suppresses AMOT expression in a PY motif-dependent manner. (A to C) Extracts from HEK293T cells transfected with myc-tagged WWOX and HA-tagged AMOT WT (A), Flag-tagged AMOT PY1, PY2, and PY3 (B), or PY123 (C) were immunoprecipitated with either normal mouse IgG or anti-myc antibody. WWOX, AMOT WT, and PY motif mutants were detected in the precipitates by WB. Expression controls for AMOT WT and PY mutants WWOX and β-actin are shown in the left panels. * indicates the IgG heavy chain. (D and E) HEK293T cells were transfected with AMOT WT or PY123 (D), PY1, PY2, and PY3 (E) mutants plus vector alone (−) or increasing amounts of WWOX, the indicated proteins were detected by WB, and AMOT levels were quantified using NIH ImageJ as shown in parentheses. (F) Quantification of the AMOT levels in panels D and E from three independent experiments. Statistical significance was analyzed by a one-way ANOVA. ns, not significant; *, P < 0.05; **, P < 0.01; ****, P < 0.0001. (G) Schematic diagram of AMOTp130 with key domains highlighted. CC, coiled-coil domain; PDZ, PSD-95/Dlg1/ZO-1 domain.