Journal: PLoS ONE

Article Title: Plasmodium sporozoites can invade hepatocytic cells independently of the Ephrin receptor A2

doi: 10.1371/journal.pone.0200032

Figure Lengend Snippet: A. Hepa1-6 cells were transfected with siRNA oligonucleotides targeting mouse EphA2 or CD81, or electroporated without siRNA (mock). Cells were infected with PyGFP sporozoites 48 hours after siRNA transfection, and the number of EEFs 24 hours post-infection was determined by fluorescence microscopy after UIS4 staining. NS, non-significant; *, p<0.05 (Kruskal-Wallis with Dunns post-test). B. HepG2/CD81cells were transfected with siRNA oligonucleotides targeting human EphA2 or human CD81, or with a control siRNA (siCtrl). Cells were infected with PyGFP sporozoites 48 hours after siRNA transfection, and the percentage of infected (GFP-positive) cells was determined by FACS 24 hours post-infection. NS, non-significant; *, p<0.05 (Kruskal-Wallis with Dunns post-test). C. HepG2/CD81 cells were transfected with siRNA targeting EphA2 or with a control siRNA (siCtrl) and infected 48 hours later with PyGFP sporozoites. Infected cultures were fixed 24 hours post-infection and stained with anti-UIS4 antibodies (red) and the nuclear stain Hoechst 33342 (blue). The images show PyGFP EEFs (green) surrounded by a UIS4-positive PVM (red). Scale bar, 10 μm. D. Hepa1-6 cells were incubated with PyGFP sporozoites in the presence of increasing concentrations of the EphA2-blocking antibody D4A2, of a control antibody (DA1E, IgG Ctrl, used at the same dilution as D4A2 highest concentration) or without antibody (Ctrl). The number of EEFs was determined 24 hours post-infection by fluorescence microscopy after UIS4 staining. NS, non-significant (Kruskal-Wallis with Dunns post-test).

Article Snippet: A control monoclonal rabbit IgG (DA1E #3900, Cell Signaling Technology) was used as a control.

Techniques: Transfection, Infection, Fluorescence, Microscopy, Staining, Incubation, Blocking Assay, Concentration Assay