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. 2014 Feb 4;136(8):3264–3270. doi: 10.1021/ja412934t

Figure 4.

Figure 4

(A) IDUP in cell lysates expressing a SNAP-tagged target protein. DNA was rapidly amplified in samples corresponding to known interactions: (B) SNAP/CAII+(GL)CBS (ΔCT = 5–6), (C) SNAP/Bcl-xL+Bad or Bak (ΔCT = 8–9), (F) SNAP-FRB+rapamycin (ΔCT = 6), and (G) FKBP-SNAP+rapamycin (ΔCT = 6). (D) A sequence corresponding to CAII+CBS was enriched ∼100-fold in a sample expressing either SNAP-CAII (lane 7) or CAII-SNAP (lane 8). (E) In samples expressing SNAP-Bcl-xL, a sequence corresponding to the interaction between Bcl-xL+Bak was enriched ∼100-fold (lane 11), but no enrichment was observed for a sequence corresponding to BclxL+BakL78A, a weakly binding mutant of the Bak peptide (lane 9). (H) Overexpression of FKBP with SNAP-FRB increased the enrichment of a sequence encoding FRB+rapamycin by 10-fold compared to a sample transfected with SNAP-FRB alone. (I) Overexpression of FRB with FKBP-SNAP also increased the enrichment of a sequence corresponding to FKBP+rapamycin by 10-fold compared to a sample transfected with FKBP-SNAP alone.