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Characterization of a highly glycosylated form of the human cytomegalovirus HLA class I homologue gpUL18, by C. Griffin, E. C. Y. Wang, B. P. McSharry, C. Rickards, H. Browne, G. W. G. Wilkinson and P. Tomasec

Journal of General Virology vol. 86, part 11, pp. 2999 - 3008

Supplementary Fig. S1.
Relative mobility of highly glycosylated forms of UL18. HFFFs were infected with HCMV AD169 or RAd-UL18 and analysed by Western blotting with anti-UL18 mAb. UL18 detected during productive HCMV infection had slower mobility during SDS-PAGE.

Supplementary Fig. S2
Amino acid sequence alignment of UL18-encoding sequences from various HCMV strains. The UL18-encoding gene from strain Toledo and clinical isolates Merlin, 6397 and 3157 were sequenced, and the translated sequences were aligned with the prototype strain AD169 sequence (GenBank accession no. P08560) using Megalign (DNASTAR). Differences to the AD169 strain UL18-encoding sequence in the other strains are boxed in grey. The GenBank accession no. for clinical isolate 6397 is bankit736177, and for clinical isolate 3157 is bankit748724. The UL18 sequences of strain Toledo (AAS48934) and Merlin (YP_081477) were identical to those determined and deposited independently in GenBank, thus the existing accession numbers are cited here.

Supplementary Fig. S3
Amino acid sequence alignment of UL18 from various HCMV strains. The DNA sequences of UL18 genes for the GenBank-deposited strain sequences Towne (accession no. AC146851), FIX (AC146907), PH (AC146904) and TR (AC146906) were translated and aligned using Megalign (DNASTAR). Differences to the AD169 UL18 sequence (P08560) are boxed in grey.

Supplementary Fig. S4
Timecourse of gpUL18 surface expression in HCMV-infected cells analysed by flow cytometry. HFFFs were mock infected or infected with HCMV AD169 or DUL18, and stained with the gpUL18 mAb 10C7, at the times indicated. Infection with AD169 UL18 deletion mutant (DUL18) or staining with mouse IgG were used as controls. Using the DUL18 mutant as a correct control, at no time after infection was it possible to detect gpUL18 on the surface of HCMV-infected cells.

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