Fig. 5: Viral ss- and dsRNA species, but not viral mRNA, are essential to trigger the translocation XRN1-DCPs into vRC. | Cell Death & Differentiation

Fig. 5: Viral ss- and dsRNA species, but not viral mRNA, are essential to trigger the translocation XRN1-DCPs into vRC.

From: Spatio-temporal characterization of the antiviral activity of the XRN1-DCP1/2 aggregation against cytoplasmic RNA viruses to prevent cell death

Fig. 5

a U-2 OS EGFP-AGO1 and mRFP-DCP1a cells were either mock treated or NDV infected (MOI = 1.0) for 1 h. Cells were then exposed to UV (500 millisevert) or CHX (100 μg/mL). Cells were harvested for either (i) RT-qPCR to quantify NDV N mRNA, or (ii) immunostaining to visualize DCP1a foci. (iii) Percentage of cells with DCP1a foci was quantified. b gRNA from respective viruses was purified, followed by transfection into (i) WT and Rig-i−/−Mda5−/− MEFs for Ifnb1 mRNA measurement by RT-qPCR, or (ii–iii) HeLa cells for immunostaining of endogenous XRN1 and quantitation of percentage of cells with XRN1 foci. c CEFs were either mock treated or infected with NDV and CVB4 (MOI = 1.0) for indicated time point. ss- and dsRNA were fractionated and analyzed on (i) agarose gel containing ethidium bromide; (ii) 7.5% PAGE by immunoblotting with anti-dsRNA antibody. d Fractionated viral ss- and dsRNA were transfected into HeLa cells and immunostained with indicated antibodies. e Percentage of cells with XRN1 aggregates was quantified. f (i) I.V.T viral mRNA was transfected into HeLa cells, followed by immunostaining for endogenous XRN1 and DCP1. (ii) Percentage of cells with XRN1-DCP1a foci was quantified. Nuclei were stained with DAPI (blue). All the white scale bars correspond to 10 μm. n.d., not detected.

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