297692.fig.002a
(a)
297692.fig.002b
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297692.fig.002c
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297692.fig.002d
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Figure 2: Purification and analysis of recombinant NHR2 containing polypeptides. (a) Schematic representation of the constructs used in this study. TAT, protein transduction domain of HIV-1 TAT protein; NLS, nuclear localization signal of SV40. (b) Purification of the recombinant NHR2 containing polypeptide TN122 by nickel affinity chromatography. SDS-PAGE and coomassie brilliant blue staining of the bacterial lysate (load), the flow through (fth), the washing fraction (w), and the eluate (e). The arrow indicates the purified TN122 protein. (c) CD spectroscopy of the NHR2 protein (NHR2-mH) at 25°C in PBS reveals an α-helical structure. (d) Glutaraldehyde crosslinking of the NHR2 protein for the indicated incubation times at room temperature and subsequent SDS-PAGE analysis. The arrows indicate the different oligomerization states of the crosslinked NHR2 proteins.