Figure 2
From: Identification and characterization of a novel Sso7d scaffold-based binder against Notch1
Characterization of Sso7d Notch1 binders. (A) Cell surface binding of candidate hits. MCF7 and MDA-MB-231 cells were stained with 1 μM candidate hits and labeled with anti-Myc-488 antibody. Red arrows indicate N9. (B) Notch1 knockdown decreases N9 binding. MCF7 cells, either transfected with siNotch1 or scramble control were stained with 1 uM N9 followed by Anti-Myc-488 antibody. Notice the shift of fluorescence peak intensity from scrambled control (blue) to siNotch 1 (green). (C) Notch1 over expression increases N9 binding. 3T3 wild type or 3T3-hNotch1 cells were incubated with 50 nM N9 and labeled with anti-Myc-488 antibody. Note the increase in N9 binding from ~5.6% in 3T3 wild type to 40.4% in 3T3-hNotch1 cells. (D) Direct binding assay of N9 and Notch1 ELR 11–15. ELR 11–15 coated wells were incubated with indicated concentrations of N9 and probed with anti-Myc-HRP antibody. Kd estimate of 0.1 ± 0.01 μM.
