Blocking experiments were used to study further the role of LDLR on DENV-2 infection described in a previous report [6 (link)]. Vero cell monolayers in six-well plates were incubated with 5 μg/well of chicken anti-LDLR antibody (Millipore, Billerica, MA, USA) at 37 °C for one hour. Following a wash, the treated cells were infected with approximately 200 PFU of DENV-2 for one hour, and a plaque assay was carried out as described above.
We also treated Vero cell monolayers in six-well plates with recombinant human LDLR at 200 ng/mL (R&D systems, Inc., Minneapolis, MN, USA) and bLF at 200 µg/mL along or together for one hour at 37 °C. After a PBS wash, the bLF and rLDLR-treated cells were infected with 200 PFU of DENV-2 in order to study the attenuation of rLDLR on anti-DENV-2 activity of bLF. The antiviral effects of the treatments were estimated in a protocol similar to the plaque reduction assay.
We also treated Vero cell monolayers in six-well plates with recombinant human LDLR at 200 ng/mL (R&D systems, Inc., Minneapolis, MN, USA) and bLF at 200 µg/mL along or together for one hour at 37 °C. After a PBS wash, the bLF and rLDLR-treated cells were infected with 200 PFU of DENV-2 in order to study the attenuation of rLDLR on anti-DENV-2 activity of bLF. The antiviral effects of the treatments were estimated in a protocol similar to the plaque reduction assay.