M. tuberculosis and SR-B1 Figure 2. Characterization of BCG and Mtb binding to SR-B1 transfected cells. R6F cells stably transfected with the indicated constructs were incubated with either BCG-lux (left panels) or Mtb-lux (right panels), and quantified for binding by measuring luciferase activity. Experiments were performed in triplicate and normalised to cell number by CFSE staining. (A) Quantification of mycobacterial binding to SR-B1 expressing cells (white bars) or vector control cells (black bars) in the presence or absence of serum, as indicated. Shown is the x-fold increase of luciferase activity compared to vector control which was set as 1. (B) Mycobacteria (BCG-lux or Mtb-lux) were pre-incubated in medium with or without 10% serum, prior binding to R6F-SR-B1 cells also in the presence or absence of 10% serum. Shown is % of luciferase activity relative to binding to R6F-SR-B1 cells in the absence of serum, which was set at 100%. (C) Effect of LDL on binding of mycobacteria to R6F cells expressing SR-B1 or SIGNR1, as indicated. Shown is % of luciferase activity relative to R6F-SR-B1 or R6F-SIGNR1 cells, respectively, in the absence of additives. (D) Effect of BAL fluid and serum (as a control) on binding of BCG-lux or Mtb-lux to SR-B1 expressing R6F cells. Shown is % of luciferase activity relative to R6F-SR-B1 in the absence of additives. (E) Effect of anti-SR-B1 antibodies on binding of BCG-lux or Mtb-lux to SR-B1 or SIGNR1 transfected R6F cells. The white bars show % of luciferase activity relative to binding in the absence of antibody (black bars). (F) SR-B1 expressing R6F cells were incubated with mycobacteria in the presence of increasing concentrations of MbCD. Binding of BCG-lux is shown as % of luciferase activity relative to control (no MbCD). doi:10.1371/journal.pone.0008448.g002 PLoS ONE | www.plosone.org 4 December 2009 | Volume 4 | Issue 12 | e8448