C-terminal domain of α-TAT1 is essential for the regulation of its acetyltransferase activity. (A) Schematic representation of GFP-α-TAT1WT (1-421 aa), GFP-α-TAT1DC (1-211 aa) and GFP-α-TAT1DN (190-421 aa). (B) HEK293T cells were transfected with the indicated plasmids, and Co-IP was performed with anti-mouse IgG and anti-Flag antibodies. (C) Co-IP assay shows the binding affinities of Flag-CK2α and GFP-α-TAT1 by overexpression of GFP-α-TAT1DC and GFP-α-TAT1DN. (D) α-TAT1 KO MEFs were transfected with GFP-C1, GFP-α-TAT1WT, GFP-α-TAT1DC, and GFP-α-TAT1DN and treated with TGF-β and bleb-bistatin. Lysates from cells were subjected to western blotting with the indicated antibodies. (E) WT MEFs were overexpressed with Myc-empty vector or α-TAT1DN-Myc and treated with blebbistatin and/or TGF-β. Next, the cells were fixed and immunostained with anti-Myc and anti-acetylated α-tubulin anti-bodies. The number of cells having dendrites whose lengths were more or less than two-times the lengths of their cell bodies was counted. For each value, measurements were made on 50 randomly photographed cells. White arrows indicate the long tail-like dendrites. Scale bar, 50 μm.