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. Author manuscript; available in PMC: 2021 Aug 25.
Published in final edited form as: Cell Rep. 2021 Aug 10;36(6):109514. doi: 10.1016/j.celrep.2021.109514

Figure 6. Nef downregulates CD8-ICL4 fusion protein.

Figure 6.

(A) CD8-ICL4 fusion proteins with an internal HA tag were created by swapping the CD8α CT with SERINC5 ICL4 and introducing substitutions into ICL4. They were expressed in HEK293T cells and detected by WB using an anti-HA antibody. SP, signal peptide; ECD, extracellular domain; TM, transmembrane; CT, cytoplasmic tail.

(B) Expression of SERINC5 proteins with the indicated substitutions was compared by WB using an anti-FLAG antibody.

(C) The indicated CD8-ICL4 fusion proteins were expressed in HEK293T cells and their expression on the cell surface was detected by flow cytometry after staining with a fluorescent anti-HA antibody.

(D) MFI values for the CD8-positive cell populations in (C) were statistically analyzed and presented.

(E) The indicated CD8-ICL4 fusion proteins were expressed with WT or ΔNef HIV-1 in HEK293T cells and analyzed as in (C).

(F) MFI values for the CD8-positive cell populations in (E) were statistically analyzed and presented.

Error bars in (D) and (F) indicate SEM calculated from three independent experiments. Statistical analysis: *p < 0.05; ns, p > 0.05.