2.8. Immunofluorescence

Immunofluorescent staining was performed as previously described (Alcendor, 2017). Briefly, chamber slide cultures containing mock infected human podocytes, podocytes infected with ZIKV and podocytes infected ZIKV after 24 h pre-treatment with DWK-1. Cells were washed twice with PBS pH 7.4, air dried, and fixed in absolute methanol for 20 min at – 20 °C. Cells were air dried for 10 min, hydrated in Tris buffered saline (pH 7.6) for 10 min, and incubated for 1 h with the 4G-2 Flavivirus group antigen monoclonal antibodies from Millipore (Temecula, CA, USA) at a dilution 1:100 in PBS pH 7.4. (Wilkerson et al., 2015).