Journal: Cells

Article Title: Host SNARE Proteins Mediate Lysosome and PVM Fusion to Support Plasmodium Liver Infection

doi: 10.3390/cells15070584

Figure Lengend Snippet: Localization of host SNARE proteins at the Plasmodium PVM during liver-stage infection. ( A – C ) Confocal live cell imaging of GFP-tagged SNARE proteins VAMP7-GFP ( A ), VAMP8-GFP ( B ), and Stx7-GFP ( C ) in P. berghei -infected HeLa cells expressing mCherry (parasite cytoplasm, red) at 6, 24, and 48 hpi. SNARE localization is shown in green. ( D – G ) Immunofluorescence analysis of Vti1B ( D ), VAMP7 ( E ), VAMP8 ( F ), and Stx7 ( G ) in HeLa cells fixed at 0.5, 1, and 1.5 hpi. SNAREs were detected with anti-GFP (green) or anti-Vti1B (green); the PVM was labeled with anti-UIS4 (red); and DAPI (blue) stained nuclei. ( H ) Localization of SNARE proteins (anti-Vti1B/anti-GFP in green) with the PVM marker UIS4 (red) at 24 hpi, visualized by expansion microscopy (5-fold expanded). Merged channels (yellow) highlight PVM-SNARE protein association. Nuclei were counterstained with DAPI (blue). Scale bars: 5 μm ( A – G ); 10 μm ( H ).

Article Snippet: The cells were then incubated in 10% FCS/PBS for 1 h, at room temperature with primary antibodies: Vti1B (mouse mAb 1:1000 (1:500 PS-ExM), BD Transduction Laboratories #611404, Allschwil, Switzerland), UIS4 (rabbit 1:1000, P. sinnis ; chicken 1:10000, Proteogenix, Schiltigheim, France), hLAMP1 (mouse mAb 1:1000 (1:500 PS-ExM), DSHB H4A3 (Iowa City, USA); rabbit pAb 1:1000, Cell Signaling #9091, Allschwill, Switzerland), GFP (rabbit pAb 1:1000, Origene TA100030, Herford, Germany; mouse mAb 1:1000, Roche AQ160, Basel, Switzerland), and α-tubulin (guinea pig pAb 1:500, ABCD AA345, Geneva, Switzerland).

Techniques: Infection, Live Cell Imaging, Expressing, Immunofluorescence, Labeling, Staining, Marker, Microscopy