DBR1 deficiency increases susceptibility to viral infection in vitro and in vivo. (A) Alignment of the sequences of segments of the DBR1 protein between the species indicated. (B) Body weights of WT (n = 10) and Dbr1Y17H/Y17H (n = 12) mice at 4–5 mo. Statistical analysis was performed with t tests. ****, P < 0.0001. Graphs depict the mean with SD and points represent biological replicates. (C) Comparison of body weight changes in WT (n = 6) and Dbr1Y17H/Y17H (n = 6) mice from birth. Graphs depict points that represent biological replicates. (D) Lariat analysis of WT and Dbr1Y17H/Y17H MEF following RNAseq. Data shown are representative of two independent experiments. (E and F) RT-qPCR assessment of WT (n = 8) and Dbr1Y17H/Y17H (n = 8) RNA lariats expression in different mouse tissues. Data representative of three independent experiments. Graphs depict mean with SD, and points represent biological replicates. (G) After the treatment of WT and Dbr1Y17H/Y17H MEFs with MG132 and CQ for 6 h, G3BP2 was detected by WB. GAPDH was used as a loading control. Data shown are representative of three independent experiments. (H and I) WT and Dbr1Y17H/Y17H MEFs were infected with viruses (VSV-GFP at an MOI of 0.1, or HSV-1-GFP at an MOI of 0.5) for 24 h; FACS analysis was then performed for the analysis of viral GFP levels (H) and RT-qPCR was used to detect expression of the indicated genes (I). Data representative of three independent experiments. Graphs depict mean with SD and points represent biological replicates. Statistical analysis was performed with one-way ANOVA with Dunnett’s multiple comparisons test. ****, P < 0.0001. (J) WB analysis after the stimulation of MEFs with polyI:C (2 µg/ml). Tubulin was used as a loading control. Data shown are representative of three independent experiments. (K) WT and Dbr1Y17H/Y17H mice were inoculated with HSV-1 (3.6 × 105 PFU/g body weight) by injection into the tail vein, and RT-qPCR was performed to assess viral replication and ISG expression in the brainstem. Data representative of three independent experiments. Graphs depict the mean with SD and points represent biological replicates. Graph points represent mouse numbers. Statistical analysis was performed with t tests. ****, P < 0.0001. (L) RT-qPCR assessment of viral gene expression in various mouse tissues. Graph points represent mouse numbers. Data representative of three independent experiments. Graphs depict the mean with SD and points represent biological replicates. Statistical analysis was performed with t tests. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Source data are available for this figure: SourceData FS5.
Figure S5.

DBR1 deficiency increases susceptibility to viral infection in vitro and in vivo. (A) Alignment of the sequences of segments of the DBR1 protein between the species indicated. (B) Body weights of WT (n = 10) and Dbr1Y17H/Y17H (n = 12) mice at 4–5 mo. Statistical analysis was performed with t tests. ****, P < 0.0001. Graphs depict the mean with SD and points represent biological replicates. (C) Comparison of body weight changes in WT (n = 6) and Dbr1Y17H/Y17H (n = 6) mice from birth. Graphs depict points that represent biological replicates. (D) Lariat analysis of WT and Dbr1Y17H/Y17H MEF following RNAseq. Data shown are representative of two independent experiments. (E and F) RT-qPCR assessment of WT (n = 8) and Dbr1Y17H/Y17H (n = 8) RNA lariats expression in different mouse tissues. Data representative of three independent experiments. Graphs depict mean with SD, and points represent biological replicates. (G) After the treatment of WT and Dbr1Y17H/Y17H MEFs with MG132 and CQ for 6 h, G3BP2 was detected by WB. GAPDH was used as a loading control. Data shown are representative of three independent experiments. (H and I) WT and Dbr1Y17H/Y17H MEFs were infected with viruses (VSV-GFP at an MOI of 0.1, or HSV-1-GFP at an MOI of 0.5) for 24 h; FACS analysis was then performed for the analysis of viral GFP levels (H) and RT-qPCR was used to detect expression of the indicated genes (I). Data representative of three independent experiments. Graphs depict mean with SD and points represent biological replicates. Statistical analysis was performed with one-way ANOVA with Dunnett’s multiple comparisons test. ****, P < 0.0001. (J) WB analysis after the stimulation of MEFs with polyI:C (2 µg/ml). Tubulin was used as a loading control. Data shown are representative of three independent experiments. (K) WT and Dbr1Y17H/Y17H mice were inoculated with HSV-1 (3.6 × 105 PFU/g body weight) by injection into the tail vein, and RT-qPCR was performed to assess viral replication and ISG expression in the brainstem. Data representative of three independent experiments. Graphs depict the mean with SD and points represent biological replicates. Graph points represent mouse numbers. Statistical analysis was performed with t tests. ****, P < 0.0001. (L) RT-qPCR assessment of viral gene expression in various mouse tissues. Graph points represent mouse numbers. Data representative of three independent experiments. Graphs depict the mean with SD and points represent biological replicates. Statistical analysis was performed with t tests. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Source data are available for this figure: SourceData FS5.

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