reference number atcc vr 118tm  (ATCC)

Journal: PLOS Pathogens

Article Title: Seoul orthohantavirus evades innate immune activation by reservoir endothelial cells

doi: 10.1371/journal.ppat.1012728

Figure Lengend Snippet: A) RLMVEC were infected with either SEOV or HTNV (MOI 0.05) or mock infected (-) and lysates were collected every 24 hours for six days (D1, D2, etc.) post-infection. Cell lysates were subjected to SDS-PAGE and immunoblot analysis for ISG expression. B) RLMVEC were infected with either SEOV or HTNV (MOI 0.05) and harvested 48 hours post-infection. Gene expression was assayed by comparative RT-PCR and averaged data from technical replicates and three experimental biological replicates are shown. Student’s T test, * denotes p<0.05. C) RLMVEC were infected with either HTNV or UV-inactivated HTNV or SEOV (MOI 0.05). Lysates were harvested at the indicated times post-infection and subjected to immunoblot analysis. D) RLMVEC were infected with either RLMVEC-passaged SEOV (RP SEOV) (MOI 0.025), UV-inactivated RLMVEC-passaged SEOV (UV RP SEOV) (0.025), or HTNV (MOI 0.025) and lysates were harvested at the indicated times post-infection and subjected to immunoblot analysis. Densitometry quantification of immunoblots in . All data shown are representative of ≥3 independent experiments.

Article Snippet: Seoul virus strain SR11 and Hantaan virus strain 76–118 were propagated on Vero E6 cells (ATCC, CRL-1586) for 12 days.

Techniques: Infection, SDS Page, Western Blot, Expressing, Gene Expression, Reverse Transcription Polymerase Chain Reaction

Journal: PLOS Pathogens

Article Title: Seoul orthohantavirus evades innate immune activation by reservoir endothelial cells

doi: 10.1371/journal.ppat.1012728

Figure Lengend Snippet: A) RLMVECs were mock-infected or infected with SEOV (MOI 0.1) for 72 hours, followed by transfection with poly(I:C) (100 ng). RNA was collected 24-hours post-treatment to assess ISG expression. B-C) RLMVECs were mock-infected or infected with SEOV (MOI 0.1) for 48 hours (B) or 72 hours (C), followed by IFNβ treatment (100 U/mL). RNA was collected at 24 hours post-treatment to evaluate ISG induction. D) RLMVECs were mock-infected or infected with SEOV (MOI 0.05) for 5 days, followed by either mock infection or superinfection with SeV (150 HAU/mL). RNA was collected 24 hours after SeV infection to measure viral genome copies and ISG expression using qRT-PCR and comparative RT-PCR, respectively. E) RLMVEC Cas9 scramble, RIG-I -/- , or MDA5 -/- cells were either mock-infected or infected with SEOV (MOI 0.05) 48 hours. Cells were the either subsequently superinfected with HTNV (MOI 0.05) or mock-infected. Lysates were collected at the indicated times post-HTNV infection and subjected to SDS-PAGE and immunoblot analysis. Densitometry quantification of immunoblots in . All data shown represent ≥3 independent experiments, with technical replicates averaged within each experiment ±SD. One-way ANOVA with multiple comparisons to mock-infected poly(I:C), IFNβ, or SeV-treated control (* denotes p<0.05).

Article Snippet: Seoul virus strain SR11 and Hantaan virus strain 76–118 were propagated on Vero E6 cells (ATCC, CRL-1586) for 12 days.

Techniques: Infection, Transfection, Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, SDS Page, Western Blot, Control

Journal: PLOS Pathogens

Article Title: Seoul orthohantavirus evades innate immune activation by reservoir endothelial cells

doi: 10.1371/journal.ppat.1012728

Figure Lengend Snippet: RLMVEC were either mock-infected or infected with SEOV (MOI 0.05) for 48 hours. Cells were subsequently either mock-infected or superinfected with HTNV (0.05 MOI) and fixed 48 hours post-HTNV infection. Images of immunostaining for SEOV nucleocapsid (green), MX1/2/3 (red), and DAPI (blue). Images were captured on an EVOS FL Auto imaging system using 20x magnification. Arrows indicate individual cells expressing both Mx1/2/3 and SEOV N. Images are representative of 2 independent experiments.

Article Snippet: Seoul virus strain SR11 and Hantaan virus strain 76–118 were propagated on Vero E6 cells (ATCC, CRL-1586) for 12 days.

Techniques: Infection, Immunostaining, Imaging, Expressing

Journal: PLOS Pathogens

Article Title: Seoul orthohantavirus evades innate immune activation by reservoir endothelial cells

doi: 10.1371/journal.ppat.1012728

Figure Lengend Snippet: Primer sequences or catalog numbers for real-time PCR analyses.

Article Snippet: Seoul virus strain SR11 and Hantaan virus strain 76–118 were propagated on Vero E6 cells (ATCC, CRL-1586) for 12 days.

Techniques: Real-time Polymerase Chain Reaction, Sequencing, Gene Expression