Journal: bioRxiv

Article Title: Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

doi: 10.1101/2023.12.08.570783

Figure Lengend Snippet: IGF2BP2-HA or VCP-HA stably expressed or not in Huh7.5 are infected or not with H/PF/2013 at an MOI of 10. Two days post infection the cells are fixed with PFA 4%. immunofluorescence-staining are performed to stain NS3 in green, dsRNA in blue and IGF2BP2-HA in red, on infected and non-infected cells.

Article Snippet: Rat polyclonal antibodies targeting DENV2 16681 NS3 which are cross-reactive with ZIKV NS3 were generated at Medimabs, Montréal, Canada) and already reported ( ).

Techniques: Stable Transfection, Infection, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

doi: 10.1101/2023.12.08.570783

Figure Lengend Snippet: FISH with IGF2BP2 and NS3 co-immunostaining was performed in Huh 7.5 after 2 days post infection with ZIKV at an MOI of 10. White arrows show the triple colocalization.

Article Snippet: Rat polyclonal antibodies targeting DENV2 16681 NS3 which are cross-reactive with ZIKV NS3 were generated at Medimabs, Montréal, Canada) and already reported ( ).

Techniques: Immunostaining, Infection

Journal: bioRxiv

Article Title: Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

doi: 10.1101/2023.12.08.570783

Figure Lengend Snippet: Huh7.5 are infected with ZIKV H/PF/2013 with an MOI of 10. Two days post infection the cells are fixed with PFA 4%. (B) immunofluorescence-staining are performed to stain NS3 in red, dsRNA in blue and (A) IGF2BP1, (B) IGF2BP3 or (C) YBX1 in green, on infected and non-infected cells.

Article Snippet: Rat polyclonal antibodies targeting DENV2 16681 NS3 which are cross-reactive with ZIKV NS3 were generated at Medimabs, Montréal, Canada) and already reported ( ).

Techniques: Infection, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Zika virus remodels and hijacks IGF2BP2 ribonucleoprotein complex to promote viral replication organelle biogenesis

doi: 10.1101/2023.12.08.570783

Figure Lengend Snippet: (A) Schematic representation of the pIRO system. Upon transfection in cells expressing the T7 RNA polymerase, this plasmid allows the cytoplasmic transcription of NS1-NS5 polyprotein under the control of T7 promoter, in a ZIKV replication-independent manner. NS1-5 polyprotein synthesis is under the control of ECMV IRES. The presence of both ZIKV 3’ NTR and 5’ cyclization sequence (5’ CS) is required for efficient VP induction. Finally, the activity of HDV ribozyme ensures that the 3’ terminus of the RNA is similar to that of vRNA genome. Huh7-Lunet-T7 were transduced with shRNA-expressing lentiviruses at a MOI of 5-10. Two days later, transduced cells were transfected with pIRO-Z plasmid. Sixteen hours later, cells were analyzed for (B) IGF2BP2 mRNA levels by RT-qPCR to measure knockdown efficiency, (C-D) transfection efficiency by confocal imaging of NS3-labeled cells, and (E) for VP content by transmission electron microscopy. Electron micrographs were used to measure (F) the percentage of cells with VPs and (G) the diameter of VPs in each condition.

Article Snippet: Rat polyclonal antibodies targeting DENV2 16681 NS3 which are cross-reactive with ZIKV NS3 were generated at Medimabs, Montréal, Canada) and already reported ( ).

Techniques: Transfection, Expressing, Plasmid Preparation, Control, Sequencing, Activity Assay, Transduction, shRNA, Quantitative RT-PCR, Knockdown, Imaging, Labeling, Transmission Assay, Electron Microscopy