Fig. 2.

Generation and purification of TAT recombinant transcription factors. a Expression of recombinant transcription factors in E. coli. Liquid culture was induced at OD₆₀₀ ~0.5 by 1 mM IPTG. The whole cell lysate was analyzed by 10% SDS-PAGE and stained with Coomassie Blue. Induction by IPTG produced an extra band (arrowed) in lane 2, 4, 6, 8 compared with lane 1, 3, 5, 7, respectively. Lane M, Pre-stained protein ladder; lane 1, 3, 5, 7, 0 h culture before IPTG induction; lane 2, 4, 6, 8, induced TAT-KLF4, TAT-c-MYC, TAT-OCT4, TAT-SOX2 fusion protein expression. b Western blot analysis of recombinant proteins whole cell lysate was separated by SDS-PAGE and analyzed by Western blot. Primary antibody: His Probe (H-15) antibody, sc-803. Lane 1, 3, 5, 7, 0 h culture before IPTG induction. Lane 2, the bigger band was TAT-Klf4, the lower one was the degraded band. Lane 4, TAT-c-Myc; Lane 6, TAT-Oct4; Lane 8, TAT-Sox2. c Analysis of purified fusion proteins. Proteins were analyzed by 10% SDS-PAGE and stained with Coomassie Blue. Lane M, Pre-stained protein ladder; Lane 1, purified TAT-Klf4; Lane 2, purified TAT-c-Myc; Lane 3, purified TAT-Oct4; Lane 4, purified TAT-Sox2