Figure 5. DARPin expression does not significantly alter expression of LmrCD proteins.

(A, B) A V5-tag was introduced in frame at the 5′-end of genomic lmrD in L. lactis (denoted L. lactis NZ9000 lmrD_V5). Plasmid-encoded DARPin activators or the control DARPin E3_5* were expressed in L. lactis NZ9000 lmrD_V5 in the presence and absence of daunomycin (14 µM for DARPin_Act3 and E3_5* and 28 µM for DARPin_Act1 and DARPin_Act2, respectively). The expression levels of genomic LmrD_V5 were then quantified by comparing the Western blot signal obtained using an anti-V5 antibody (A) with total protein detected by SYPRO ruby staining (B). (C) The relative amounts of LmrD_V5 expression were quantified by densitometry. Each bar represents the average of three independent data points (n = 3) of which one data point is shown in (A) and (B).