Skip to main content
. 2012 Nov 30;7(11):e50396. doi: 10.1371/journal.pone.0050396

Figure 3. Stable expression of sh1CD81 downregulates surface CD81 expression and increases TCR mediated activation.

Figure 3

(a) GFP expression before and after puromycin selection in VL3.3M2 cell lines transfected with pLMP encoding CD81 shRNA (sh1CD81) shows the percentage of cells expressing CD81 shRNA. Shaded histograms show fluorescensce of untransfected VL3.3M2 cells and black lined histograms show GFP expression in transfected and antibiotic selected cells. (b) CD81 shRNAs decrease surface CD81 expression in stably transfected VL3.3M2 cells. Relative MFI of CD81 expression on the surface of VL3.3M2 cells that are untransfected (U), or stably transfected with empty pLMP constructs (LMP) or with pLMP-sh1CD81 constructs (sh1CD81) or single cell cloned stable sh1CD81 expressing clones (clone1 and clone2) was determined by flow cytometry and plotted as bar graphs. Surface CD81 expression of untransfected VL3.3M2 cells was set to 100. (c) Relative MFI of surface CD69 expression on VL3.3M2 cells stimulated with plate bound anti-TCRß (1 µg) and anti-CD4 (1 µg). Surface CD69 expression of empty pLMP transfected VL3.3M2 cells was set to 100.