Journal: bioRxiv
Article Title: CD164 is an endolysomal host factor for entry of Clade A New World Arenaviruses
doi: 10.64898/2026.04.21.719929
Figure Lengend Snippet: (A) Coomassie-stained SDS-PAGE of soluble proteins used in . 2.5 µg of PNGaseF treated, nondenatured, or denatured purified protein. (B) Nonlinear fit of binding curves at pH 5.0 (blue) and 7.4 (grey) for PICV sGP1, PARV sGP1, FLEXV sGP1, and MACV sGP1 to 100 nM of immobilized sCD164. Curves represent the averaged final response at the end of association (n=3, independent experiments in triplicate). (C) Binding of 5 µM MACV sGP1 (maroon triangle) or 1 µg mouse anti-human TfR1 (CD71) antibody (cyan diamond) to 100 nM of immobilized hTfR1-Fc as a function of pH. Normalized to max binding at pH 8.0 (n=2, independent experiments in duplicate). (D) pH 7.4 binding curve of MACV sGP1 to hTfR1 (100 nM). Left: 1:1 Langmuir fit; middle: nonlinear fit; right: Scatchard plot. Binding affinity (K D ) calculated from 1:1 Langmuir fit, K D = 2.52 µM ± 0.006 µM. (n=2, independent experiments in duplicate). (E) Representative histogram of mean fluorescence intensity (MFI) from . Cells bound with 2.5 µM AF647-labeled PICV sGP1 at pH=6.0 (orange: CD164 KO +WT-CD164 PM ; purple: CD164 KO ) or pH 7.4 (red: CD164 KO +CD164 PM ; blue: CD164 KO ) (n=3, independent experiments in triplicate) (F) MFI (/10,000) of CD164 KO +WT-CD164 PM or CD164 KO cells from and S4E, labeled with 5 µM AF647-PICV sGP1 at pH 6.0 and then washed at pH 7.4 (hashed) or 6.0 (solid). (n=3, independent experiments in triplicate).
Article Snippet: For each binding assay, sCD164-Fc, hTfR1-Fc, sCRD-Fc, or mutant CD164, were first immobilized on GatorBio anti-human IgG Fc Gen II (HFCII) probes (GatorBio,160024) for 300 sec.
Techniques: Staining, SDS Page, Purification, Binding Assay, Fluorescence, Labeling