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. Author manuscript; available in PMC: 2025 Apr 18.
Published in final edited form as: Cell Chem Biol. 2024 Mar 19;31(4):658–668.e14. doi: 10.1016/j.chembiol.2024.02.004

Figure 3. Nef PROTAC treatment restores cell surface CD4 and MHC-I expression in T cells.

Figure 3.

The human T cell line CEM-T4 was engineered to express a Nef-eGFP fusion protein under the control of a doxycycline (Dox) inducible promoter. In the absence of Dox, these cells express endogenous CD4 and MHC-I on their surface; addition of Dox induces Nef-eGFP expression which leads to receptor downregulation. A) Representative flow cytometry result with Nef PROTACs FC-14369 (active) and FC-14379 (inactive). B) Active Nef PROTACs from the NanoBRET ubiquitination assay were screened for cell surface receptor rescue in triplicate. Bar height indicates the mean value ± SE; individual data points are also shown. The structures of the analogs with little to no activity in this assay (FC-13890, FC-14373, FC-14379, FC-14388) are shown in the Supplemental Information, Figure S2.