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. Author manuscript; available in PMC: 2012 Sep 15.
Published in final edited form as: J Immunol. 2012 Feb 15;188(6):2794–2804. doi: 10.4049/jimmunol.1102068

Figure 5. UL40 promotes cell surface expression of gpUL18 and HLA-E.

Figure 5

(A) Cell surface expression of gpUL18, MHC-I, gB, gH and gN compared on HFFF cells infected with HCMV strain AD169 or AD169ΔUL40 for 72h (30 PFU/cell). (B) Cell surface expression of gpUL18 on cells infected with HCMV strain AD169, AD169ΔUL40, AD169ΔUL18 (30 PFU/cell) for 72h. (C) gpUL18 surface expression on cells co-infected with RAd-UL18 (100 PFU/cell) and either RAd-UL40 or the vector control (RAd-CTRL) (30 PFU/cell) for 72h. (D) HFFF infected with RAd-UL18 at the multiplicities of infection (m.o.i) indicated and RAd-UL40 or RAd-CTRL (30 PFU/cell) for 72h. Results are means ± SEM of duplicate samples (Two-Way ANOVA test with Bonferroni post-tests). (E) Cell surface expression of HLA-E (antibody 3D12) and MHC-I on HFFF infected with strain AD169ΔUL16, AD169ΔUL18 or AD169ΔUL40 (15 PFU/cell) for 96h (F) Western blot of HFFF infected with AD169ΔUL16, AD169ΔUL18 or AD169ΔUL40 (15 PFU/cell) for 96h. (G) gpUL18 detected by western blot in HFFF infected with RAd-UL18 (100 PFU/cell) and either RAd-UL40 or RAd-CTRL (30 PFU/cell) for 72h. Cell extracts were digested with Endo H [E] or PNGase F [P] or no enzyme [Mock; M]. Endo H-resistant gpUL18 glycoform (Inline graphic), Endo H-sensitive gpUL18 glycoform before (Inline graphic) and after (Inline graphic) digestion, PNGase-digested gpUL18 forms (Inline graphic) are indicated. (H) Following cell surface biotinylation and immunoprecipitation, gpUL18, HLA-E, HLA-HC and CD155 cell surface expression were compared on western blot in cells infected with AD169ΔUL16, AD169ΔUL18 or AD169ΔUL40 (15 PFU/cell) for 96h.