Fig. S1. Recombinant IsdB and IsdB Y165A/H166E/Y167A binding to human haemoglobin (hHb). rIsdB was immobilized in and incubated with different concentration of hHb. Bound hHb was detected with polyclonal rabbit anti-hHb serum followed by HRP-conjugated goat anti-rabbit IgG. Values represent the mean of triplicate wells. Binding assays was preformed three times with similar results.


Fig. S2. Inducible expression of IsdB by L. lactis. Whole-cell immunoblot to detect surface expression of IsdB by L. lactis NZ9000. L. lactis pNZ8037isdB and L. lactis pNZ8037 were grown to exponential phase and induced with nisin (at indicated concentrations). After 4 h post induction the bacteria were harvested, dotted on cellulose membranes (at indicted concentrations) and subjected to immunoblotting. Detection was performed using rabbit anti-IsdB IgG followed by HRP-conjugated goat anti-rabbit IgG.


Fig. S3. Real-time measurement of attachment and spreading of breast cancer MCF-7 cells. (A) Fibronectin (Fn) was immobilized in microtitre wells, blocked with BSA and incubated with equal numbers of MCF-7 cells supplemented with either IsdA, IsdH-GST, GST or IsdB. XCELLigence was used to measure the real-time impedance in those wells. Cells attachment and spreading led to increased impedance which corresponded to higher cell index (CI) values. (A) Traces of CI changes representing MCF-7 spreading on Fn in the presence of IsdA (light grey), IsdH-GST (middle grey), GST (dark grey) or IsdB (black) over time are shown. (B) Slope changes (slope h−1) of CI values representing MCF-7 cells attachment to fibronectin upon incubation with different proteins (as indicated). Slope changes were calculated for CI traces within time points of 30 min to 240 min.


Table S1. Primers used in this study.

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