Fig. 3.

Mapping the nuclear localization domain of Myb2. The sequence of Myb2 is depicted as the N terminus, R2R3 DNA-binding domain, and C terminus. Series deletions from the N and C termini of Myb2 were performed, and the mutant proteins are referred to as ΔN or ΔC, respectively, with a number to show the deletion boundary. (B) Subcellular localization of 4HA-Myb2, Δ48, Δ55, Δ143, and Δ132 was detected by IFA with a mouse anti-HA antibody. The signal was shown as green fluorescence (FITC). The nucleus was stained with DAPI, and cell morphology was recorded by phase-contrast microscopy. Bar, 5 μm. (C) Total lysates (TL) from cells overexpressing 4HA-Myb2 and Δ48, Δ55, Δ143, and Δ132 were fractionated into the nuclear (N) and cytosolic (C) fractions. Protein samples were separated in 12% gel for Western blotting using a rat anti-HA antibody. Duplicate blots were examined by the anti-acetyl-histone H3(Lys9) (α-H3K9-Ac) and anti-CypA1 (α-CypA1) antibodies.