Fig. 3
From: Pho dynamically interacts with Spt5 to facilitate transcriptional switches at the hsp70 locus
Pho interacts dynamically with Spt5 and dSfmbt. a Multiple protein alignment between Pho and YY1, the mouse and human homologue of Pho. The REPO domain is surrounded by a blue rectangle, while the Zn-finger domains are surrounded by an orange rectangle. The asterisk mark on the bottom line ‘consensus’ denotes the amino acid residues that are fully conserved between the three species. b Representation of the N- and C-termini of Pho and the mutant constructs generated for this study. c, d co-IP assays of S2 DRSC cells transiently transfected with plasmids expressing FLAG-tagged dSfmbt or Spt5 (FLAG-dSfmbt, FLAG-Spt5) and HA-tagged Pho, PhoV164D, Pho ΔREPO, EGFP (HA-Pho, HA-PhoV164D, HA-Pho REPO del, HA-EGFP) as indicated. Cell lysates were used for pull-downs using an anti-FLAG antibody and were later probed by Western blot using an anti-HA antibody. MW = molecular weight in kDa. e co-IP assays of S2 DRSC cells transiently transfected with plasmids expressing FLAG-tagged dSfmbt or Spt5 (FLAG-dSfmbt, FLAG-Spt5) and HA-tagged Pho or Spt4 (HA-Pho, HA-Spt4) as indicated. S2 DRSC cells were either maintained at normal growth temperature (25 °C) or heat shocked at 37 °C for 15 min, cell lysates were used for pull-downs using an anti-FLAG antibody and were later probed by Western blot using an anti-HA antibody. MW = molecular weight in kDa