FIG 2.

Interference with cell surface heparan sulfate either by enzymatic removal or by blockage via anti-3-OS HS (G2) peptide negatively affects HCMV infection of HIS cells. (A) Cultured HIS cells were either mock treated or untreated (Un) with serum-free medium or pretreated with 1 mg/ml control peptide (Cp; RVCGSIGKEVLG), anti-3-OS HS (G2; MPRRRRIRRRQK) peptide, and heparinase I (H+) (10 IU) for 2 h followed by addition of the reporter HCMV at an MOI of 4 to individual wells for 12 h before the cells were washed to remove the unbound virus. The cells were kept at 37°C in 5% CO₂ for 9 days before the effect of treatment was assessed. As indicated in panel A, HCMV entry was assessed via ONPG substrate (3.0 mg/ml) for quantitation of β-galactosidase activity expressed from the input viral genome. The enzymatic activity was measured at the optical density at 410 nm (OD₄₁₀). Asterisks indicate significant difference from other treatments (P < 0.05, t test), and error bars represent SD (n = 4). (B) X-Gal staining of the HIS cells for blue plaques or foci in response to following treatments: mock treated (a), Cp treated (b), heparinase treated H+ [c]), and anti-3-OS HS (G2) peptide treated (d).