Figure 5

(A) Model structure of human ubiquitin protein labelled with doubly Maleimide DOTA Gadolinium tag in the S20C and G35C position. (B) Derived bulk intracellular concentration for the Ub-ma-DOTA-Gd³⁺ delivered cells. The concentration was estimated interpolating the Integral value of the in-cell spectra Gd³⁺ transition with a calibration curve. (C) Acquired EDFS spectra for the HEK 293 T and HeLa cells over the center of the resonator dip under overcoupling conditions. (D) DEER spectra acquired for the in vitro (blue traces) and the relative in-cell sample (red traces).