reovirus 3  (ATCC)


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    ATCC reovirus 3
    We have demonstrated DRACO efficacy against a broad spectrum of viruses.
    Reovirus 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Broad-Spectrum Antiviral Therapeutics"

    Article Title: Broad-Spectrum Antiviral Therapeutics

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0022572

    We have demonstrated DRACO efficacy against a broad spectrum of viruses.
    Figure Legend Snippet: We have demonstrated DRACO efficacy against a broad spectrum of viruses.

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    reovirus t3d stock vr 824  (ATCC)


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    ATCC reovirus t3d stock vr 824
    Reovirus T3d Stock Vr 824, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    reovirus 3  (ATCC)


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    ATCC reovirus 3
    We have demonstrated DRACO efficacy against a broad spectrum of viruses.
    Reovirus 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Broad-Spectrum Antiviral Therapeutics"

    Article Title: Broad-Spectrum Antiviral Therapeutics

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0022572

    We have demonstrated DRACO efficacy against a broad spectrum of viruses.
    Figure Legend Snippet: We have demonstrated DRACO efficacy against a broad spectrum of viruses.

    Techniques Used:

    heterogeneous reovirus t3d stock  (ATCC)


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    ATCC heterogeneous reovirus t3d stock
    Heterogeneous Reovirus T3d Stock, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    heterogenous t3d stock  (ATCC)


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    ATCC heterogenous t3d stock
    Heterogenous T3d Stock, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    reovirus 3  (ATCC)


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    ATCC reovirus 3
    Reovirus 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    reovirus 3  (ATCC)


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    ATCC reovirus 3
    Reovirus 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    reovirus 3  (ATCC)


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    ATCC reovirus 3
    Reovirus 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mammalian reovirus serotype dearing 3  (ATCC)


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    ATCC mammalian reovirus serotype dearing 3
    Mammalian Reovirus Serotype Dearing 3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    reovirus t3d strain  (ATCC)


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    ATCC reovirus t3d strain
    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus <t>T3D</t> strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Reovirus T3d Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "DHX15 is required to control RNA virus-induced intestinal inflammation"

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    Journal: Cell reports

    doi: 10.1016/j.celrep.2021.109205

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Techniques Used: Enzyme-linked Immunosorbent Assay, shRNA, Infection, Expressing

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Expressing

    (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).

    Techniques Used: Injection, Enzyme-linked Immunosorbent Assay, Plaque Assay, Flow Cytometry, Infection, Staining

    (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.
    Figure Legend Snippet: (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.

    Techniques Used: Infection, Purification, Incubation, Immunoprecipitation, Transfection, Plasmid Preparation

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Knock-Out, Transgenic Assay, shRNA, Software

    reovirus t3d strain  (ATCC)


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    ATCC reovirus t3d strain
    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus <t>T3D</t> strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Reovirus T3d Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 1 article reviews
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    reovirus t3d strain - by Bioz Stars, 2024-05
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    1) Product Images from "DHX15 is required to control RNA virus-induced intestinal inflammation"

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    Journal: Cell reports

    doi: 10.1016/j.celrep.2021.109205

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Techniques Used: Enzyme-linked Immunosorbent Assay, shRNA, Infection, Expressing

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Expressing

    (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).
    Figure Legend Snippet: (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).

    Techniques Used: Injection, Enzyme-linked Immunosorbent Assay, Plaque Assay, Flow Cytometry, Infection, Staining

    (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.
    Figure Legend Snippet: (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.

    Techniques Used: Infection, Purification, Incubation, Immunoprecipitation, Transfection, Plasmid Preparation

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Knock-Out, Transgenic Assay, shRNA, Software

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    ATCC reovirus t3d strain
    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus <t>T3D</t> strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).
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    We have demonstrated DRACO efficacy against a broad spectrum of viruses.

    Journal: PLoS ONE

    Article Title: Broad-Spectrum Antiviral Therapeutics

    doi: 10.1371/journal.pone.0022572

    Figure Lengend Snippet: We have demonstrated DRACO efficacy against a broad spectrum of viruses.

    Article Snippet: Dengue type 2 (New Guinea C, VR-1584), Amapari (VR-477), Tacaribe (VR-1272), Guama (Be An 277, VR-407; Be Ar 12590, VR-420), murine adenovirus (VR-550), Theiler's murine encephalomyelitis (VR-57), reovirus 3 (VR-824), influenza H1N1 A/PR/8/34 (ATCC VR-1469), influenza H1N1 A/WS/33 (ATCC VR-1520), rhinovirus 1B (VR-481), rhinovirus 2 (VR-482), rhinovirus 14 (VR-284), and rhinovirus 30 (VR-505) were obtained from ATCC.

    Techniques:

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Journal: Cell reports

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    doi: 10.1016/j.celrep.2021.109205

    Figure Lengend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H), and IL-18 (C, F, and I) production from human HT-29 IECs with the indicated shRNA after a 20-h infection with enteric RNA viruses, including simian rotavirus SA-11 strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a multiplicity of infection (MOI) of 10. Mock, scrambled shRNA (sh-Ctrl)-treated human HT-29 IECs without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in human HT-29 IECs infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Article Snippet: Reovirus T3D strain , ATCC , Cat#VR-824.

    Techniques: Enzyme-linked Immunosorbent Assay, shRNA, Infection, Expressing

    (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Journal: Cell reports

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    doi: 10.1016/j.celrep.2021.109205

    Figure Lengend Snippet: (A–I) ELISA of IFN-β (A, D, and G), IFN-λ3 (B, E, and H) and IL-18 (C, F, and I) production in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice after a 20-h infection with enteric RNA viruses, including rotavirus EW strain (A–C) and reovirus T3D strain (D–F), or DNA virus HSV-1 KOS strain (G–I) at a MOI of 10. Mock, cells without virus infection. Each circle represents an individual independent experiment, and small solid black lines indicate the average of triplicates. (J–L) Quantification of expression of rotavirus NSP5 gene (J), reovirus S4 gene (K), and HSV-1 VP16 gene (L) relative to GAPDH in mouse primary IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected by rotavirus (J), reovirus (K), or HSV-1(L) as in (A)–(I). Data are represented as means ± SEMs. NS, p > 0.05, ***p < 0.001 (unpaired t test).

    Article Snippet: Reovirus T3D strain , ATCC , Cat#VR-824.

    Techniques: Enzyme-linked Immunosorbent Assay, Infection, Expressing

    (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).

    Journal: Cell reports

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    doi: 10.1016/j.celrep.2021.109205

    Figure Lengend Snippet: (A) Survival of 5-week-old wild-type Dhx15 fl/fl and Dhx15 IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 10 8 plaque-forming units [PFUs] per mouse). (B–D) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 10 8 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA. (E and F) The wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 10 8 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs. (G) Flow cytometry analysis of CD4 + T cells, CD8 + T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies. (H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G). (I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice as in (E). Scale bars represent 200 μm. (J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).

    Article Snippet: Reovirus T3D strain , ATCC , Cat#VR-824.

    Techniques: Injection, Enzyme-linked Immunosorbent Assay, Plaque Assay, Flow Cytometry, Infection, Staining

    (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.

    Journal: Cell reports

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    doi: 10.1016/j.celrep.2021.109205

    Figure Lengend Snippet: (A) IB analysis of endogenous proteins of DHX15 and NLRP6 precipitated with anti-NLRP6 from whole-cell lysates of IECs from wild-type Dhx15 fl/fl and Dhx15 IEC-KO mice infected without (Mock) or with reovirus (Reo) T3D strain at MOI of 5. (B) Schematic diagram showing full-length (Full) DHX15 and its truncations with deletion of various domains (left margin); numbers at ends indicate amino acid positions (top). DEXDc, DEAD-like helicases superfamily domain; HELICc, helicase superfamily c-terminal domain; HA2, helicase-associated domain 2; DUF, domain of unknown function. (C) IB analysis of purified FLAG-tagged NLRP6 with anti-FLAG (bottom blot), and IB analysis (with anti-HA) of purified HA-tagged full-length DHX15 (Full) and DHX15 truncation mutants alone (top blot) including N-terminal domain (N), DEAD-like helicases superfamily domain (DEXDc), and C-terminal domain (C), or after incubation with FLAG-tagged NLRP6 and immunoprecipitation with anti-FLAG (center blot). (D) IB analysis of DHX15, NLRP6, caspase-1, and ASC in HEK293T cells transfected with Myc-ASC and co-transfected with or without FLAG-caspase-1, FLAG-NLRP6, HA-DHX15 (Full for full-length, or C for its truncate containing C-terminal domain), or HA-vector control followed by immunoprecipitation with anti-Myc antibody. (E) IB analysis of DHX15, NLRP6, caspase-1, ASC, full-length IL-18, and its cleaved IL-18 in HEK293T cells transfected with HA-IL-18, Myc-ASC, and FLAG-caspase-1, and co-transfected with or without FLAG-NLRP6, HA-DHX15, or HA-vector control with different doses as indicated. The position of protein markers (shown in kDa) is indicated at right.

    Article Snippet: Reovirus T3D strain , ATCC , Cat#VR-824.

    Techniques: Infection, Purification, Incubation, Immunoprecipitation, Transfection, Plasmid Preparation

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: DHX15 is required to control RNA virus-induced intestinal inflammation

    doi: 10.1016/j.celrep.2021.109205

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Reovirus T3D strain , ATCC , Cat#VR-824.

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Knock-Out, Transgenic Assay, shRNA, Software