Fig. 3

a Truncation of TTC fragment from Bcl2-TTC. The 254 N-terminal residues of TTC were genetically deleted by introducing an additional XhoI restriction site, such that the resulting Bcl2-hTTC fusion protein consisted of the 6 × His tag sequence, Bcl-2 and the C-terminal 204 residues of TTC. The estimated molecular weight of Bcl2-hTTC was 53 kDa, which is almost identical to that of full-length TTC (55 kDa). b Purification and detection of TTC and Bcl2-hTTC. Total protein (0.025 μg and 0.10 μg for TTC and Bcl2-hTTC, respectively) was loaded onto the gel. Purified fusion proteins were stained with CBB, and were further detected using an anti-6 × His antibody and anti-Bcl-2 antibody. c Cell surface binding of fusion proteins. Neuronal binding capacity of Bcl2-hTTC to differentiated PC12 cells as detected by an anti-Bcl-2 antibody and anti-6 × His antibody. Bcl2-hTTC showed neuronal binding activity that was similar to that of TTC (Fig. 2b) and Bcl2-TTC (Fig. 2d). Differentiated PC12 cells added vehicle alone were used as a control. Scale bar = 25 μm