Fig 1.
Percentage of diarrhea, acholic stool, and biliary obstructive disease (acholic stool, oily fur, and growth retardation) in Stat1-/- mice suckling mice orally infected with rRRV, rSA11, and mono-reassortant RVs with different NSP1 or VP4 encoding genes.
(A) rRRV; (B) rSA11; (C) rSA11 with RRV VP4; (D) rSA11 with RRV NSP1; (E) rRRV with SA11 NSP1; (F) rRRV with UK NSP1. N = 8–12 in each group. (G) Daily weight changes in Stat1-/- suckling mice orally infected with mock, rRRV, rSA11 and reciprocal NSP1 reassortants on indicated days following RV infection. * (P<0.05, rRRV or rSA11-RRV NSP1 vs. rSA11 or rRRV-SA11 NSP1). Without indication, P>0.05, not significant.
Fig 2.
Percentage of diarrhea, acholic stool, and biliary obstructive disease (acholic stool, oily fur, and growth retardation) in Stat1-/- mice suckling mice orally infected with recombinant RVs encoding different regions of RRV NSP1.
(A) Western blot images of IRF3 degradation in MA104 cells (multiplicity of infection of 3, 8 hours post infection). (B) rSA11 with RRV NSP1; (C) rSA11 with RRV NSP1 deletion; (D) rSA11 with RRV NSP1 16 amino acids truncated at the C-terminal end; (E) daily weight changes; (F) Kaplan–Meier survival curves. Del: deletion.
Fig 3.
Fecal virus shedding and levels of viral replication in liver and pancreas in Stat1-/- suckling mice orally infected with rRRV, rSA11, and mono-reassortant RVs with different NSP1 or VP4 encoding genes.
(A) Fecal virus shedding; (B) levels of viral replication in livers; (C) levels of viral replication in pancreases. Brackets show statistically significant difference between the viral strains and groups. * (P<0.05), ** (P<0.01). n.t., not detected.
Fig 4.
Immunofluorescent staining of liver, bile duct, and pancreas of Stat1-/- suckling mice orally infected with mock, rRRV, rSA11 and reciprocal NSP1 mono-reassortants at day 10 post infection.
Tissues were stained with Alexa Fluor 488 labeled mouse monoclonal antibody against RV VP6 (green), Alexa Fluor 594 labeled rabbit monoclonal antibodies against α-SMA or CK19, and DAPI (blue). (A) liver; (B) biliary duct; (C) pancreas. Scale bar: 1 mm.
Fig 5.
H&E staining of liver in Stat1-/- suckling mice orally infected with rRRV, rSA11 and reciprocal NSP1 mono-reassortants at day 10 post infection.
(A) uninfected mouse; (B) rRRV; (C) rSA11; (D) rSA11 with RRV NSP1; (E) rRRV with SA11 NSP1. Red arrows indicate areas of inflammation in region of portal triads. Scale bar: 200 μm.
Fig 6.
H&E staining of extra-hepatic biliary ducts in Stat1-/- suckling mice orally infected with rRRV, rSA11 and reciprocal NSP1 mono-reassortants at day 10 post infection.
(A) uninfected mouse; (B) rRRV; (C) rSA11; (D) rSA11 with RRV NSP1; (E) rRRV with SA11 NSP1. Black arrows indicate open bile ducts and red arrows indicate areas of severe biliary obstruction. Scale bar: 200 μm.
Fig 7.
H&E staining of pancreas in Stat1-/- suckling mice orally infected with rRRV, rSA11 or reciprocal NSP1 mono-reassortants at day 10 post infection.
(A) uninfected mouse; (B) rRRV; (C) rSA11; (D) rSA11 with RRV NSP1; (E) rRRV with SA11 NSP1. Black circles indicate healthy pancreatic parenchyma and white circles indicate focal areas of inflammation. Scale bar: 200 μm.
Fig 8.
Blood chemistry analysis in serum of Stat1-/- suckling mice orally infected with rRRV, rSA11 and reciprocal NSP1 mono-reassortants.
(A) serum conjugated bilirubin; (B) serum lipase; (C) serum amylase; (D) serum alanine aminotransferase (ALT). Brackets show statistically significant differences between combined groups with RRV NSP1 vs SA11 NSP1. * (P<0.05).