Skip to main content
. 2015 Apr 29;89(14):7202–7213. doi: 10.1128/JVI.00854-15

FIG 1.

FIG 1

Specific binding of HKU1-S1 to RD cells. (A) SDS-PAGE of expressed recombinant S1 or control (Ctrl.) proteins. All proteins were expressed in 293T cells and purified by the use of protein A Sepharose beads. Purified proteins were run on SDS-PAGE and stained by the use of Coomassie blue. (B) FACS analysis of HKU1-S1(600)-mFc (5 μg/ml) binding to RD and HeLa cells. HKU3-323-mFc was used as a negative-control protein. (C) HKU1-S1(600)-mFc binding to a molecule(s) located on the RD cell surface. Results of immunofluorescence microscopy imaging of RD cells stained by HKU1-S1(600)-mFc or the control protein, HKU3-323-mFc, are shown. Cell membranes were stained with FM-4-64 (red), the nuclei were stained with Hoechst dye 33258 (blue), and the HKU1-S1 staining was detected by an FITC-labeled anti-mouse Fc antibody. (D) HKU1-S1(600)-mFc binding to RD cells in a dose-dependent manner in a FACS analysis. (E) FACS analysis of the N termini of HKU1-S1 and OC43-S1 binding to RD cells. HKU3-323-mFc at 10 μg/ml was used as a control (Ctrl.) in panels D and E. The graphs shown in panels B to E are representative of the results of at least two independent experiments for each panel.