Journal: iScience
Article Title: A key role for the exoribonuclease XRN1 in regulating the hepatitis B viral transcriptome
doi: 10.1016/j.isci.2026.116328
Figure Lengend Snippet: HBV induces P-bodies (A and B) Effect of HBV infection on XRN1 and EDC4 expression. Mock or HBV-infected HepG2-NTCP cells were cultured for up to 9 days (A) P-body components XRN1 and EDC4, together with HBsAg and β-actin, were detected at 3, 6, and 9 dpi by western blotting. (B) EDC4 (green) and DAPI nuclei (blue) were detected by immunofluorescence at 6 dpi, with a representative image shown (scale bars, 25 μm). The average number of EDC4 puncta relative to nuclei was quantified in 9 images from 3 independent experiments, with significance assessed using a Mann-Whitney test (∗∗∗ p < 0.001, ∗ p < 0.05). (C) Abundance of viral and host RNAs. Total HBV RNAs (HBV-total, yellow) and host Peptidylprolyl isomerase B RNA (PPIB, red) and nuclear DAPI (blue) were imaged at 6 dpi (scale bars, 20 μm). (D) Replication of in vitro transcribed HBV pgRNA. A schematic of in vitro transcribed (IVT) WT or Δε pgRNA and their viral products. Mock or IVT HBV RNA-transfected (100 ng) Huh 7.5 cells were cultured for 2, 4, or 6 days. Cells were imaged for HBc (magenta) (scale bars, 75 μm), intracellular HBV DNA, and secreted HBsAg assessed by qPCR or ELISA, respectively. (E) pCpg RNA levels after 6 days were measured by RT-qPCR, and the data were normalized to a housekeeping gene, B2M , respectively. (F) EDC4 puncta in HBV RNA-transfected cells. Mock or IVT HBV RNA-transfected (100 ng) Huh 7.5 cells (6 days) were stained for EDC4 expression and puncta number and size quantified in 15 images from 3 independent experiments. Mean EDC4 values are expressed relative to the number of nuclei per image, with statistical significance assessed using a Mann-Whitney test with correction for multiple comparisons (∗∗∗ p < 0.001 and ∗ p < 0.05).
Article Snippet: Antibodies used in this study (and their supplier) are: EDC4 (FORTIS), DDX6 (FORTIS), HBsAg (Stratech), HBcAg (Dako), HBcAg (Beacle Inc.) XRN1 (Thermo Fisher Scientific), β-Actin (Sigma), HBsAg (Abcam), anti-mouse Immunoglobulin HRP (Dako), anti-rabbit IgG HRP (Cytiva), anti-mouse IgG HRP (Abcam), anti-rabbit IgG (Alexa Fluor 488, 594 and 633, Thermo Fisher Scientific) and anti-7-methylguanosine (m 7 G)-Cap (Caltag Medsystems Ltd).
Techniques: Infection, Expressing, Cell Culture, Western Blot, Immunofluorescence, MANN-WHITNEY, In Vitro, Transfection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Staining
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