encephalomyocarditis virus emcv strain emc Search Results


emcv  (ATCC)
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ATCC emcv
a, Schematic representation of the TRIM shRNA screen in 293.rKSHV.219 cells. b, rKSHV.219 reactivation from a following lentiviral transduction of non-targeting control shRNA (sh.C) or TRIM-specific shRNAs (x axis) by analyzing RFP-positive cells using fluorescence-activated cell sorting (FACS). c, TRIM transcripts in iSLK.219 cells treated with doxycycline (1 μ g ml-1) for 3 days to induce rKSHV.219 reactivation, determined by qRT-PCR. d, Left, TRIM43 transcripts in EBV-infected AKBM or EBV-negative BJAB cells that were mock-treated or treated with 100 μ g ml-1 anti-human immunoglobulin G (IgG) for 24 h to induce EBV reactivation, determined by qRT-PCR. Right, EBV reactivation determined by analyzing BMRF1 transcripts by qRT-PCR. e, TRIM43 transcripts in human foreskin fibroblast (HFF) cells infected with HCMV (MOI of 3) for 24 h, determined by qRT-PCR. f, TRIM43 transcripts in Huh7 cells infected with VSV (MOI 0.01), DV (MOI 1) or HSV 1 (MOI 1) for 18 h, assessed by qRT-PCR. g, TRIM43 mRNA expression in BAL samples from patients with acute pulmonary infection, assessed by qRT-PCR. h, Heatmap summarizing the results from the siRNA mini-screen to test the effect of TRIM knockdown on the replication of HSV-1, Ad, <t>EMCV</t> and VSV (Supplementary Fig. 2d–g). i, Left, EBV BMRF1 transcripts in AGS-EBV cells transfected with non-targeting control siRNA (si.C), si.TRIM43 or si.TRIM4 (negative control), determined by qRT-PCR at 96 h post-transfection. Middle and right, knockdown of TRIM43 and TRIM4, determined by qRT-PCR at 48 h after siRNA transfection. j, EBV Zta-protein abundance of AGS-EBV cells from i, determined by immunoblot (IB) using anti-Zta at 120 h after siRNA transfection. k, HCMV IE-1 protein-positive HFF cells following transfection with si.TRIM43 or si.C and subsequent infection with HCMV (50 IE-1 units) for 24 h, determined by immunofluorescence (IF). Data represent mean and s.d. of n = 3 (biological replicates) (b,c–f,i,k), or mean and s.d. of 18 HSV-1-positive or 16 HSV-1-negative BAL samples (g). Statistical significance was calculated by unpaired two-tailed t-test (b,d,i,k), or two-tailed Mann-Whitney U test (g). *P < 0.05, **P < 0.01, ***P < 0.001. Exact P values for b are provided in Supplementary Table 1. Results are representative of one screen (b) or three independent experiments (c–f,h–k). nd, not detectable.
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a, Schematic representation of the TRIM shRNA screen in 293.rKSHV.219 cells. b, rKSHV.219 reactivation from a following lentiviral transduction of non-targeting control shRNA (sh.C) or TRIM-specific shRNAs (x axis) by analyzing RFP-positive cells using fluorescence-activated cell sorting (FACS). c, TRIM transcripts in iSLK.219 cells treated with doxycycline (1 μ g ml-1) for 3 days to induce rKSHV.219 reactivation, determined by qRT-PCR. d, Left, TRIM43 transcripts in EBV-infected AKBM or EBV-negative BJAB cells that were mock-treated or treated with 100 μ g ml-1 anti-human immunoglobulin G (IgG) for 24 h to induce EBV reactivation, determined by qRT-PCR. Right, EBV reactivation determined by analyzing BMRF1 transcripts by qRT-PCR. e, TRIM43 transcripts in human foreskin fibroblast (HFF) cells infected with HCMV (MOI of 3) for 24 h, determined by qRT-PCR. f, TRIM43 transcripts in Huh7 cells infected with VSV (MOI 0.01), DV (MOI 1) or HSV 1 (MOI 1) for 18 h, assessed by qRT-PCR. g, TRIM43 mRNA expression in BAL samples from patients with acute pulmonary infection, assessed by qRT-PCR. h, Heatmap summarizing the results from the siRNA mini-screen to test the effect of TRIM knockdown on the replication of HSV-1, Ad, EMCV and VSV (Supplementary Fig. 2d–g). i, Left, EBV BMRF1 transcripts in AGS-EBV cells transfected with non-targeting control siRNA (si.C), si.TRIM43 or si.TRIM4 (negative control), determined by qRT-PCR at 96 h post-transfection. Middle and right, knockdown of TRIM43 and TRIM4, determined by qRT-PCR at 48 h after siRNA transfection. j, EBV Zta-protein abundance of AGS-EBV cells from i, determined by immunoblot (IB) using anti-Zta at 120 h after siRNA transfection. k, HCMV IE-1 protein-positive HFF cells following transfection with si.TRIM43 or si.C and subsequent infection with HCMV (50 IE-1 units) for 24 h, determined by immunofluorescence (IF). Data represent mean and s.d. of n = 3 (biological replicates) (b,c–f,i,k), or mean and s.d. of 18 HSV-1-positive or 16 HSV-1-negative BAL samples (g). Statistical significance was calculated by unpaired two-tailed t-test (b,d,i,k), or two-tailed Mann-Whitney U test (g). *P < 0.05, **P < 0.01, ***P < 0.001. Exact P values for b are provided in Supplementary Table 1. Results are representative of one screen (b) or three independent experiments (c–f,h–k). nd, not detectable.

Journal: Nature microbiology

Article Title: Centrosomal protein TRIM43 restricts herpesvirus infection by regulating nuclear lamina integrity

doi: 10.1038/s41564-018-0285-5

Figure Lengend Snippet: a, Schematic representation of the TRIM shRNA screen in 293.rKSHV.219 cells. b, rKSHV.219 reactivation from a following lentiviral transduction of non-targeting control shRNA (sh.C) or TRIM-specific shRNAs (x axis) by analyzing RFP-positive cells using fluorescence-activated cell sorting (FACS). c, TRIM transcripts in iSLK.219 cells treated with doxycycline (1 μ g ml-1) for 3 days to induce rKSHV.219 reactivation, determined by qRT-PCR. d, Left, TRIM43 transcripts in EBV-infected AKBM or EBV-negative BJAB cells that were mock-treated or treated with 100 μ g ml-1 anti-human immunoglobulin G (IgG) for 24 h to induce EBV reactivation, determined by qRT-PCR. Right, EBV reactivation determined by analyzing BMRF1 transcripts by qRT-PCR. e, TRIM43 transcripts in human foreskin fibroblast (HFF) cells infected with HCMV (MOI of 3) for 24 h, determined by qRT-PCR. f, TRIM43 transcripts in Huh7 cells infected with VSV (MOI 0.01), DV (MOI 1) or HSV 1 (MOI 1) for 18 h, assessed by qRT-PCR. g, TRIM43 mRNA expression in BAL samples from patients with acute pulmonary infection, assessed by qRT-PCR. h, Heatmap summarizing the results from the siRNA mini-screen to test the effect of TRIM knockdown on the replication of HSV-1, Ad, EMCV and VSV (Supplementary Fig. 2d–g). i, Left, EBV BMRF1 transcripts in AGS-EBV cells transfected with non-targeting control siRNA (si.C), si.TRIM43 or si.TRIM4 (negative control), determined by qRT-PCR at 96 h post-transfection. Middle and right, knockdown of TRIM43 and TRIM4, determined by qRT-PCR at 48 h after siRNA transfection. j, EBV Zta-protein abundance of AGS-EBV cells from i, determined by immunoblot (IB) using anti-Zta at 120 h after siRNA transfection. k, HCMV IE-1 protein-positive HFF cells following transfection with si.TRIM43 or si.C and subsequent infection with HCMV (50 IE-1 units) for 24 h, determined by immunofluorescence (IF). Data represent mean and s.d. of n = 3 (biological replicates) (b,c–f,i,k), or mean and s.d. of 18 HSV-1-positive or 16 HSV-1-negative BAL samples (g). Statistical significance was calculated by unpaired two-tailed t-test (b,d,i,k), or two-tailed Mann-Whitney U test (g). *P < 0.05, **P < 0.01, ***P < 0.001. Exact P values for b are provided in Supplementary Table 1. Results are representative of one screen (b) or three independent experiments (c–f,h–k). nd, not detectable.

Article Snippet: EMCV (strain EMC) was purchased from ATCC.

Techniques: shRNA, Transduction, Control, Fluorescence, FACS, Quantitative RT-PCR, Infection, Expressing, Knockdown, Transfection, Negative Control, Quantitative Proteomics, Western Blot, Immunofluorescence, Two Tailed Test, MANN-WHITNEY