Quantitative proteomics identifies 30 CD81 receptor interactions in hepatoma cells.

<p>(A) Schematic overview of the experimental setup used to define the CD81-interactome in human hepatoma cell lines. (B) Immunoblot analysis of CD81- and HA-IPs from Lunet N hCD81HA and Lunet N hCD81 cells using anti-CD81 or anti-HA antibodies, as indicated. GAPDH served as loading control. L = lysate, FT = flow through, E = eluate. Representative of 4 biological replicates. (C) LFQ intensities of proteins in CD81- and HA-IPs from the indicated cell line. CD81 (green) and SCARB1 (black) served as positive and APOL2 (white) as negative control. CAPN5 (red) was discovered as CD81 interactor in hepatoma cells. Median of 4 biological replicates. (D) Number of proteins significantly enriched in the indicated co-IPs and membrane associated fraction. (E) Volcano plot visualizing two-sample t-test comparing LFQ intensities of proteins found in CD81-IPs from Lunet N hCD81 and Lunet N. For each protein the t-test difference (log₁₀) of CD81 versus control co-IP of 4 biological replicates is plotted against the p value (-log₁₀). FDR = 0.01; s0 = 2. Proteins significantly enriched are highlighted in dark grey. CD81 (green), SCARB1 (black) APOL2 (white) and CAPN5 (red) are highlighted. (F) Overlap of significantly enriched proteins found in anti-CD81co-IPs from Lunet N hCD81 and Lunet N hCD81HA. See also <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007111#ppat.1007111.s001" target="_blank">S1 Fig</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007111#ppat.1007111.s002" target="_blank">S2 Fig</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007111#ppat.1007111.s010" target="_blank">S1 Table</a> and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007111#ppat.1007111.s011" target="_blank">S2 Table</a>.