FIG 2.

Regulation of galectin 9 expression by HCMV (A) Parental HF, V-HF, or nPro-HF were infected at an MOI of 3 with HCMV or UV-HCMV before cell lysates were harvested at 48 h p.i. and immunoblotted for Gal-9 (R&D Systems) and GAPDH (Santa Cruz Biotechnology). (B) Media from mock-, HCMV-, or UV-HCMV-infected parental HFs were harvested at 24 h p.i. and filtered through a 0.1-μm-pore-size filter before being added to fresh HF, V-HF, or nPro-HF monolayers and incubated for 48 h before cell lysates were immunoblotted for Gal-9 and GAPDH. Blots are representative of at least 3 independent biological replicates. (C) HFs were mock, HCMV, or UV-HCMV infected in the presence of an IFN-β-blocking Ab or an isotype control. Cell lysates were harvested at 48 h p.i. before immunoblotting for Gal-9 and GAPDH. (D) HFs were treated with media from mock-, HCMV-, or UV-HCMV-infected cultures (harvested at 24 h p.i. and filtered through a 0.1-μm-pore-size filter) in the presence of an IFN-β-blocking Ab or an isotype control. Cell lysates were generated at 48 h postinfection/treatment before immunoblotting for Gal-9 and GAPDH. (E) HFs were treated with recombinant IFN-β at the concentrations indicated before immunoblotting for Gal-9 and GAPDH at 48 h posttreatment. HFs were also mock and HCMV infected in parallel as indicated. Blots are representative of at least 3 independent biological replicates.