short interfering rna sirna Search Results


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  • 99
    Thermo Fisher sirna
    Sirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher lipofectamine 2000
    Lipofectamine 2000, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipofectamine 2000/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lipofectamine 2000 - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    98
    Thermo Fisher sirna transfection
    Sirna Transfection, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna transfection/product/Thermo Fisher
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna transfection - by Bioz Stars, 2021-05
    98/100 stars
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    86
    Horizon Discovery sirna
    Localization of MX2 and NUP214 following transfection with Nup/NTR targeting <t>siRNA</t> continued. Deconvolution microscopic images (single optical sections) of <t>HeLa</t> cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP214 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Representative of two independent experiments, with at least three images acquired per experiment.
    Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna/product/Horizon Discovery
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sirna - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    N/A
    Target species mouse Gene Silencers generally consist of pools of three to five target specific 19 25 nucleotide sequences in length For independent verification of DUXBL gene silencing results individual
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    N/A
    Target species mouse Gene Silencers generally consist of pools of three to five target specific 19 25 nucleotide sequences in length For independent verification of Rhox2a gene silencing results individual
      Buy from Supplier

    N/A
    Target species mouse Gene Silencers generally consist of pools of three to five target specific 19 25 nucleotide sequences in length For independent verification of Rhox2b gene silencing results individual
      Buy from Supplier

    N/A
    Target species mouse Gene Silencers generally consist of pools of three to five target specific 19 25 nucleotide sequences in length For independent verification of Rhox3a gene silencing results individual
      Buy from Supplier

    Image Search Results


    Localization of MX2 and NUP214 following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP214 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2 and NUP214 following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP214 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Effect of Nup and NTR depletion on primate lentivirus infection and MX2 sensitivity. Infectivity of (A) HIV-2 or (B) SIVmac GFP reporter virus in HeLa cells stably transduced with doxycycline-inducible MX2 in the presence (open circles) or absence (filled circles) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on primate lentivirus infection and MX2 sensitivity. Infectivity of (A) HIV-2 or (B) SIVmac GFP reporter virus in HeLa cells stably transduced with doxycycline-inducible MX2 in the presence (open circles) or absence (filled circles) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Infection, Stable Transfection, Transduction, Transfection

    Localization of MX2 and NUP214 following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP214 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2 and NUP214 following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP214 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Effect of Nup and NTR depletion on HIV-1 infection and MX2 sensitivity in the presence of CsA. ( A–B ) Infectivity of HIV-1 GFP reporter virus in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the absence of doxycycline and in the presence (asterisks) or absence (circles, HeLa or triangles, HT1080) of CsA. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). ( C–D ) Infectivity of HIV-1 GFP reporter virus in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence of CsA and in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on HIV-1 infection and MX2 sensitivity in the presence of CsA. ( A–B ) Infectivity of HIV-1 GFP reporter virus in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the absence of doxycycline and in the presence (asterisks) or absence (circles, HeLa or triangles, HT1080) of CsA. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). ( C–D ) Infectivity of HIV-1 GFP reporter virus in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence of CsA and in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Infection, Stable Transfection, Transduction, Transfection

    Localization of CPSF6 and NUP98 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of CPSF6 and NUP98 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Localization of SV40 NLS-GFP-LacZ following transfection with Nup/NTR targeting siRNA continued. HeLa cells stably transduced with SV40 NLS-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of SV40 NLS-GFP-LacZ following transfection with Nup/NTR targeting siRNA continued. HeLa cells stably transduced with SV40 NLS-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Stable Transfection, Transduction, Staining

    Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Localization of MX2 and NUP62 following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP62 (green), and Hoechst-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2 and NUP62 following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP62 (green), and Hoechst-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Knockdown of Nups and NTRs in HOS cells. Western blot analysis of Nup and NTR expression levels in A) dividing and B) non-dividing (aphidicolin treated) HOS cells 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each blot correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although blotting experiments carried out in triplicate (approximately 5), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Knockdown of Nups and NTRs in HOS cells. Western blot analysis of Nup and NTR expression levels in A) dividing and B) non-dividing (aphidicolin treated) HOS cells 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each blot correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although blotting experiments carried out in triplicate (approximately 5), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Western Blot, Expressing, Transfection, Generated, Infection

    Localization of MX2(N25)-GFP-LacZ following transfection with Nup/NTR targeting siRNA. HeLa cells stably transduced with MX2(N25)-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2(N25)-GFP-LacZ following transfection with Nup/NTR targeting siRNA. HeLa cells stably transduced with MX2(N25)-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Stable Transfection, Transduction, Staining

    Effect of Nup and NTR depletion on HIV-1 CA mutant infection and MX2 sensitivity. Infectivity of HIV-1 CA mutant GFP reporter viruses in HOS cells or HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on HIV-1 CA mutant infection and MX2 sensitivity. Infectivity of HIV-1 CA mutant GFP reporter viruses in HOS cells or HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Mutagenesis, Infection, Stable Transfection, Transduction, Transfection

    Localization of CPSF6 and NUP98 in HeLa cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of CPSF6 and NUP98 in HeLa cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Localization of MX2(N25)-GFP-LacZ following transfection with Nup/NTR targeting siRNA continued. HeLa cells stably transduced with MX2(N25)-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2(N25)-GFP-LacZ following transfection with Nup/NTR targeting siRNA continued. HeLa cells stably transduced with MX2(N25)-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Stable Transfection, Transduction, Staining

    Knockdown of Nups and NTRs in HT1080 cells. Western blot analysis of Nup and NTR expression levels in A) dividing and B) non-dividing (aphidicolin treated) HT1080 cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. (n/a – not tested since HT1080 cells do not express NUP210, see Figure 2—figure supplement 1 ). Bands for each blot correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on the subcellular localization of MX2 and Nups, as well as on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although blotting experiments carried out in triplicate (approximately 5), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Knockdown of Nups and NTRs in HT1080 cells. Western blot analysis of Nup and NTR expression levels in A) dividing and B) non-dividing (aphidicolin treated) HT1080 cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. (n/a – not tested since HT1080 cells do not express NUP210, see Figure 2—figure supplement 1 ). Bands for each blot correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on the subcellular localization of MX2 and Nups, as well as on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although blotting experiments carried out in triplicate (approximately 5), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Western Blot, Expressing, Stable Transfection, Transduction, Transfection, Generated, Infection

    Localization of MX2, NUP153, and RANBP2 in HeLa cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2, NUP153, and RANBP2 in HeLa cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Knockdown of Nups and NTRs in non-dividing HeLa cells. Western blot analysis of Nup and NTR expression levels in growth arrested (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each Nup/NTR correspond to those indicated in Figure 2 . Each blot is generated from an experiment in which the effect of siRNA knockdown on WT HIV-1 infection was similar to that shown in . The complete array of blots was conducted once, although randomly selected blotting experiments carried out in triplicate (approximately 10), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Knockdown of Nups and NTRs in non-dividing HeLa cells. Western blot analysis of Nup and NTR expression levels in growth arrested (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each Nup/NTR correspond to those indicated in Figure 2 . Each blot is generated from an experiment in which the effect of siRNA knockdown on WT HIV-1 infection was similar to that shown in . The complete array of blots was conducted once, although randomly selected blotting experiments carried out in triplicate (approximately 10), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Western Blot, Expressing, Stable Transfection, Transduction, Transfection, Generated, Infection

    Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Cell-cycle profile of HOS cells following transfection with Nup/NTR targeting siRNA. Cell-cycle profile of HOS cells 64 hr after transfection with the indicated siRNA or following aphidicolin treatment, color coded by subcomplex as in Figure 4A . DNA content was determined by flow cytometry following staining with the DNA dye DRAQ5. For each plot, 3000–12000 events were collected. Representative of two independent experiments. 2N and 4N DNA content are indicated for the Control siRNA transfected sample.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Cell-cycle profile of HOS cells following transfection with Nup/NTR targeting siRNA. Cell-cycle profile of HOS cells 64 hr after transfection with the indicated siRNA or following aphidicolin treatment, color coded by subcomplex as in Figure 4A . DNA content was determined by flow cytometry following staining with the DNA dye DRAQ5. For each plot, 3000–12000 events were collected. Representative of two independent experiments. 2N and 4N DNA content are indicated for the Control siRNA transfected sample.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Flow Cytometry, Cytometry, Staining

    Localization of CPSF6 and NUP98 in HeLa cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of CPSF6 and NUP98 in HeLa cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HeLa cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Localization of SV40 NLS-GFP-LacZ following transfection with Nup/NTR targeting siRNA. HeLa cells stably transduced with SV40 NLS-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of SV40 NLS-GFP-LacZ following transfection with Nup/NTR targeting siRNA. HeLa cells stably transduced with SV40 NLS-GFP-LacZ fusion protein (green) and doxycycline-inducible MX2-RFP in the absence of doxycycline fixed and stained with Hoescht 64 hr after transfection with the indicated siRNA. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Stable Transfection, Transduction, Staining

    Effect of ARFAPTIN2 fusion protein expression on lentivirus and HIV-1 CA mutant infection. ( A ) Infection of HeLa cells stably transduced with doxycycline-inducible arfaptin2 myc-tagged fusion proteins with various GFP reporter viruses in the presence (white bars) or absence (black bars) of doxycycline. Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. ( B ) Infectivity of HIV-1 N57S mutant GFP reporter virus in HT1080 cells stably transduced with arfaptin2 fusion proteins in the presence (white bars) or absence (black bars) of doxycycline and the presence or absence of CsA. Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. (C) Infectivity of HIV-1-GFP reporter virus in dividing or non-dividing (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible arfaptin2 or MX2(N91)-arfaptin2 in the presence (open circles) or absence (filled circles) of doxycycline. Titers are mean ± sem, n = 3 technical replicates and are representative of three independent experiments. ( D ) Infectivity of HIV-1 GFP reporter virus in growth arrested (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible MX2(N91)-ARFAPTIN2 in the presence (open circles) or absence (filled circles) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of ARFAPTIN2 fusion protein expression on lentivirus and HIV-1 CA mutant infection. ( A ) Infection of HeLa cells stably transduced with doxycycline-inducible arfaptin2 myc-tagged fusion proteins with various GFP reporter viruses in the presence (white bars) or absence (black bars) of doxycycline. Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. ( B ) Infectivity of HIV-1 N57S mutant GFP reporter virus in HT1080 cells stably transduced with arfaptin2 fusion proteins in the presence (white bars) or absence (black bars) of doxycycline and the presence or absence of CsA. Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. (C) Infectivity of HIV-1-GFP reporter virus in dividing or non-dividing (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible arfaptin2 or MX2(N91)-arfaptin2 in the presence (open circles) or absence (filled circles) of doxycycline. Titers are mean ± sem, n = 3 technical replicates and are representative of three independent experiments. ( D ) Infectivity of HIV-1 GFP reporter virus in growth arrested (aphidicolin treated) HeLa cells stably transduced with doxycycline-inducible MX2(N91)-ARFAPTIN2 in the presence (open circles) or absence (filled circles) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Expressing, Mutagenesis, Infection, Stable Transfection, Transduction, Transfection

    Effect of Nup and NTR depletion on HIV-1 infection in HOS cells. ( A ) Infectivity of HIV-1 GFP reporter virus in dividing (top) and non-dividing (bottom) HOS cells 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ±sem, n = 3 technical replicates, representative of three independent experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on HIV-1 infection in HOS cells. ( A ) Infectivity of HIV-1 GFP reporter virus in dividing (top) and non-dividing (bottom) HOS cells 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ±sem, n = 3 technical replicates, representative of three independent experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Infection, Transfection

    Knockdown of Nups and NTRs in dividing HeLa cells. Western blot analysis of Nup and NTR expression levels in HeLa cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each Nup/NTR correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on the subcellular localization of MX2 and Nups, as well as on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although randomly selected blotting experiments carried out in triplicate (approximately 10), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Knockdown of Nups and NTRs in dividing HeLa cells. Western blot analysis of Nup and NTR expression levels in HeLa cells stably transduced with doxycycline-inducible MX2 64 hr after transfection with the indicated siRNA color coded by subcomplex as in Figure 4A . Red boxes highlight lanes for corresponding antibody-siRNA pairs. Bands for each Nup/NTR correspond to those indicated in Figure 2 . Each blot was generated from an experiment in which the effect of siRNA knockdown on the subcellular localization of MX2 and Nups, as well as on WT HIV-1 infection was similar to that shown below. The complete array of blots was conducted once, although randomly selected blotting experiments carried out in triplicate (approximately 10), as well as infectivity data indicated both consistent levels of knockdown and pleiotropic effects across experiments.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Western Blot, Expressing, Stable Transfection, Transduction, Transfection, Generated, Infection

    Localization of MX2, NUP153, and RANBP2 in HeLa cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2, NUP153, and RANBP2 in HeLa cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Efficient siRNA-mediated knockdown of Nups and pleiotropic effects of Nup depletion in HOS cells. Heat map representing protein levels of Nups or NTRs (color coded by subcomplex as in Figure 4A ) in dividing (top) and non-dividing (bottom) HOS cells 64 hr after transfection with the indicated siRNA. Protein levels are expressed as ratios of Nup/NTR expression:LAMIN B1 expression, based on the blots shown in Figure 5—figure supplement 6 , normalized to control siRNA transfected cells that were assigned a value of 1.0 (black). Reduced expression is indicated in gray-white, with no detectable expression assigned a zero value (white). Red boxes highlight corresponding antibody-siRNA pairs.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Efficient siRNA-mediated knockdown of Nups and pleiotropic effects of Nup depletion in HOS cells. Heat map representing protein levels of Nups or NTRs (color coded by subcomplex as in Figure 4A ) in dividing (top) and non-dividing (bottom) HOS cells 64 hr after transfection with the indicated siRNA. Protein levels are expressed as ratios of Nup/NTR expression:LAMIN B1 expression, based on the blots shown in Figure 5—figure supplement 6 , normalized to control siRNA transfected cells that were assigned a value of 1.0 (black). Reduced expression is indicated in gray-white, with no detectable expression assigned a zero value (white). Red boxes highlight corresponding antibody-siRNA pairs.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing

    Cell-cycle profile of HeLa cells following transfection with Nup/NTR targeting siRNA. Cell-cycle profile of HeLa cells 64 hr after transfection with the indicated siRNA or following aphidicolin treatment, color coded by subcomplex as in Figure 4A . DNA content was determined by flow cytometry following staining with the DNA dye propidium iodide (PI). For each plot, 3000–12000 events per sample were collected. Representative of two independent experiments. 2N and 4N DNA content are indicated for the Control siRNA transfected sample.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Cell-cycle profile of HeLa cells following transfection with Nup/NTR targeting siRNA. Cell-cycle profile of HeLa cells 64 hr after transfection with the indicated siRNA or following aphidicolin treatment, color coded by subcomplex as in Figure 4A . DNA content was determined by flow cytometry following staining with the DNA dye propidium iodide (PI). For each plot, 3000–12000 events per sample were collected. Representative of two independent experiments. 2N and 4N DNA content are indicated for the Control siRNA transfected sample.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Flow Cytometry, Cytometry, Staining

    Efficient siRNA-mediated knockdown of Nups and pleiotropic effects of Nup depletion in HT1080 cells. Heat map representing protein levels of Nups or NTRs (color coded by subcomplex as in Figure 4A ) in dividing (top) and non-dividing (bottom) HT1080 cells 64 hr after transfection with the indicated siRNA. Protein levels are expressed as ratios of Nup/NTR expression:LAMIN B1 expression, based on the blots shown in Figure 5—figure supplement 5 , normalized to control siRNA transfected cells that were assigned a value of 1.0 (black). Reduced expression is indicated in gray-white, with no detectable expression assigned a zero value (white). Red boxes highlight corresponding antibody-siRNA pairs.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Efficient siRNA-mediated knockdown of Nups and pleiotropic effects of Nup depletion in HT1080 cells. Heat map representing protein levels of Nups or NTRs (color coded by subcomplex as in Figure 4A ) in dividing (top) and non-dividing (bottom) HT1080 cells 64 hr after transfection with the indicated siRNA. Protein levels are expressed as ratios of Nup/NTR expression:LAMIN B1 expression, based on the blots shown in Figure 5—figure supplement 5 , normalized to control siRNA transfected cells that were assigned a value of 1.0 (black). Reduced expression is indicated in gray-white, with no detectable expression assigned a zero value (white). Red boxes highlight corresponding antibody-siRNA pairs.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing

    Localization of MX2 and NUP62 following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP62 (green), and Hoechst-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2 and NUP62 following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HeLa cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP62 (green), and Hoechst-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Effects of depleting individual Nups on NPC integrity and function, and MX2 localization. Summary of the localization of Nups, MX2-RFP, CPSF6-RFP, or NLS-GFP-LacZ fusions upon siRNA transfection of HeLa or HT1080 cells shown in Figure 6 , and Figure 6—figure supplements 1 – 18 . Aberrant localization following siRNA transfection in ≥~80% of cells is indicated by an ‘x’ and normal localization is indicated by a dot.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effects of depleting individual Nups on NPC integrity and function, and MX2 localization. Summary of the localization of Nups, MX2-RFP, CPSF6-RFP, or NLS-GFP-LacZ fusions upon siRNA transfection of HeLa or HT1080 cells shown in Figure 6 , and Figure 6—figure supplements 1 – 18 . Aberrant localization following siRNA transfection in ≥~80% of cells is indicated by an ‘x’ and normal localization is indicated by a dot.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection

    Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of MX2, NUP153, and RANBP2 in HT1080 cells following transfection with Nup/NTR targeting siRNA continued. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing MX2-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP153 (green), RANBP2 (purple), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 5 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Localization of CPSF6 and NUP98 in HT1080 cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Localization of CPSF6 and NUP98 in HT1080 cells following transfection with Nup/NTR targeting siRNA. Deconvolution microscopic images (single optical sections) of HT1080 cells expressing CPSF6-RFP (red, stably transduced with a doxycycline inducible vector), immunoflourescently stained NUP98 (green), and Hoescht-stained DNA. Cells were fixed and stained 64 hr after transfection with the indicated siRNA. Optical sections are approximately through the center of the vertical dimension on the nucleus. Scale bar = 10 μm. Representative of two independent experiments, with at least three images acquired per experiment.

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Transfection, Expressing, Stable Transfection, Transduction, Plasmid Preparation, Staining

    Effect of Nup and NTR depletion on HIV-1 A92E and N57S CA mutant infection in the presence of CsA. ( A–C ) Infectivity of HIV-1 A92E or N57S GFP reporter viruses in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the absence of doxycycline and in the presence (asterisks) or absence (circles, HeLa or triangles, HT1080) of 5 μM CsA. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). X-axis legend for A-B is below graph B, x-axis legend for C-D is below graph D. ( D ) Infectivity of HIV-1 N57S GFP reporter virus in HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence of CsA and in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on HIV-1 A92E and N57S CA mutant infection in the presence of CsA. ( A–C ) Infectivity of HIV-1 A92E or N57S GFP reporter viruses in HeLa or HT1080 cells stably transduced with doxycycline-inducible MX2 in the absence of doxycycline and in the presence (asterisks) or absence (circles, HeLa or triangles, HT1080) of 5 μM CsA. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). X-axis legend for A-B is below graph B, x-axis legend for C-D is below graph D. ( D ) Infectivity of HIV-1 N57S GFP reporter virus in HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence of CsA and in the presence (open symbols) or absence (filled symbols) of doxycycline. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Mutagenesis, Infection, Stable Transfection, Transduction, Transfection

    Effect of Nup and NTR depletion on non-primate lentivirus infection and MX2 sensitivity. Infectivity of ( A ) EIAV or ( B-D ) FIV GFP reporter virus in ( A-B ) HeLa or ( C ) HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence (open symbols) or absence (filled symbols) of doxycycline, or ( D ) HOS cells. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Journal: eLife

    Article Title: Nuclear pore heterogeneity influences HIV-1 infection and the antiviral activity of MX2

    doi: 10.7554/eLife.35738

    Figure Lengend Snippet: Effect of Nup and NTR depletion on non-primate lentivirus infection and MX2 sensitivity. Infectivity of ( A ) EIAV or ( B-D ) FIV GFP reporter virus in ( A-B ) HeLa or ( C ) HT1080 cells stably transduced with doxycycline-inducible MX2 in the presence (open symbols) or absence (filled symbols) of doxycycline, or ( D ) HOS cells. Cells were infected 64 hr after transfection with siRNA (color coded by subcomplex as in Figure 4A ). Titers are mean ± sem, n = 3 technical replicates, representative of three independent experiments. n/a – not included due to insufficient knockdown (NUP37, RAE1) or not expressed (NUP210).

    Article Snippet: RNA interference HOS, HeLa, or HT1080 cells were reverse transfected with 25 pmol of siRNA ( ; SMARTpool, Dharmacon) using Lipofectamine RNAiMax (Invitrogen) at a concentration of 3 × 104 cells/ml in 6- or 12-well plates.

    Techniques: Infection, Stable Transfection, Transduction, Transfection