Figure 6. Menin functionally interacts with LEDGF.
A. Schematic structures of menin and its mutants. The abilities of each protein to associate with MLL or LEDGF and to rescue MLL-dependent transcription are summarized on the right. *, indicates impaired association as opposed to complete loss of association. ND, not determined.
B. Various HA-tagged menin proteins were transiently expressed with (+) or without (-) (f)ME in 293T cells and subjected to IP with anti-HA antibody, followed by immunoblotting with anti-FLAG, anti-HA, and anti-LEDGF antibodies.
C. Experimental scheme for rescue of Men1 knockdown by wild type or mutant menin proteins. The time points at which CFUs or gene expression were measured are indicated.
D. Western blot analysis shows expression of menin mutants in ME-transformed cells detected by anti-HA and anti-MLLN antibodies, respectively.
E. Relative CFU are shown for ME-transformed cells transduced with various menin mutants with (+) or without (-) Men1 knock down (vector control was arbitrarily set at 100%). Error bars represent the standard deviations of three independent analyses.
F. Relative expression levels of Hoxa9 and Men1 are shown for first round colonies after Men1 knockdown. Expression levels are normalized to Gapdh, and expressed relative to the ME/vector value (arbitrarily set at 100%). Error bars represent the standard deviations of triplicate PCR analyses. P values for differences compared to wild type menin rescue were determined by unpaired t test (*, P<0.0005; **, P<0.005).