Figure 6.

The Ydj1 interaction is stronger with Ssa2 than Ssa4. (A) The His₆-Ydj1 immobilized beads were incubated with yeast lysate from A2 (1X) or A4 (3X) cells expressing FLAG-tagged v-Src. The bound proteins were probed with anti-FLAG, anti-Hsp70, anti-Hsp90, or anti-Ydj1 antibody. Relatively higher amount of Hsp70 from the A2 strain was detected indicating a stronger interaction of Ydj1 with Ssa2 than Ssa4. Panel toward right depicts quantification of respective western blot. ^#A mutant of Ydj1 [His₆-Ydj1(H34Q)] was used as negative control. (B) The biolayer interferometry sensogram showing interaction of Ydj1 with increasing concentrations of Hsp70 isoforms (Ssa2 or Ssa4) as analytes. A much stronger interaction of Ydj1 with Ssa2 than Ssa4 was observed at similar concentrations of the two Hsp70 isoforms. Inset shows the zoomed-in view of Ydj1 interaction with Ssa4. The labels are represented as the same order of magnitude of the curve. Error bars represent SD from three different biological replicates. P-values were calculated using the Student’s t-test and A2 as a control. w.r.t is defined as with respect to.