Fig. 1
From: h-Prune as a novel binding protein of NS5A that regulates ERK1/2 activation
Schematic representation of the full-length and the four different NS5A derivatives and determining the NS5A and h-prune binding site. (A) The DNA fragments of the NS5A derivatives were PCR-amplified and subcloned into pHybLex/Zeo (a). The four NS5A derivatives (designated NS5A-F, -a, -b, and -c) were tested to determine their suitability as baits for full-scale yeast two-hybrid screening in a colony-lift filter assay (b). The h-prune-binding fragments were NS5A-F and NS5A-c. pHybLex/Zeo-Fos2 and pYESTrp-Jun were cotransfected into the L40 yeast strain as a positive control [(+) control], and pHybLex/Zeo-Lamin was transfected alone for the negative control [(−) control] in the colony-lift filter assay (c), unless specified elsewhere. (B) GST pull-down assay with GST (control) and GST-NS5A (pull-down) of yeast extracts expressing h-prune with the MAL tag. NS5A directly interacted with h-prune