module Search Results


nebnext poly a mrna magnetic isolation module  (New England Biolabs)
99
New England Biolabs nebnext poly a mrna magnetic isolation module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Nebnext Poly A Mrna Magnetic Isolation Module, supplied by New England Biolabs, 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/product/module/bio_rxiv__64898__2026__03__05__709814-53-27-33?v=New+England+Biolabs
Average 99 stars, based on 1 article reviews
nebnext poly a mrna magnetic isolation module - by Bioz Stars, 2026-08
99/100 stars
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nebnext companion module  (New England Biolabs)
97
New England Biolabs nebnext companion module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Nebnext Companion Module, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pm41872458-301-12-12?v=New+England+Biolabs
Average 97 stars, based on 1 article reviews
nebnext companion module - by Bioz Stars, 2026-08
97/100 stars
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da tailing module  (New England Biolabs)
96
New England Biolabs da tailing module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Da Tailing Module, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/us11781179-199-20-25?v=New+England+Biolabs
Average 96 stars, based on 1 article reviews
da tailing module - by Bioz Stars, 2026-08
96/100 stars
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nebnext quick ligation module  (New England Biolabs)
98
New England Biolabs nebnext quick ligation module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Nebnext Quick Ligation Module, supplied by New England Biolabs, 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/product/module/pm42012196-218-76-76?v=New+England+Biolabs
Average 98 stars, based on 1 article reviews
nebnext quick ligation module - by Bioz Stars, 2026-08
98/100 stars
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nebnext ultra ii fs module  (New England Biolabs)
96
New England Biolabs nebnext ultra ii fs module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Nebnext Ultra Ii Fs Module, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pmc13099204-85-7-7?v=New+England+Biolabs
Average 96 stars, based on 1 article reviews
nebnext ultra ii fs module - by Bioz Stars, 2026-08
96/100 stars
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module  (New England Biolabs)
96
New England Biolabs module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Module, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/10__12688_slash_f1000research__10571__1-84-21-22?v=New+England+Biolabs
Average 96 stars, based on 1 article reviews
module - by Bioz Stars, 2026-08
96/100 stars
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clampfit module  (Danaher Inc)
96
Danaher Inc clampfit module
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Clampfit Module, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pmc06596023-475-8-16?v=Danaher+Inc
Average 96 stars, based on 1 article reviews
clampfit module - by Bioz Stars, 2026-08
96/100 stars
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programmable syringe pump  (New Era Pump Systems Inc)
95
New Era Pump Systems Inc programmable syringe pump
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Programmable Syringe Pump, supplied by New Era Pump Systems Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pmc09427107-270-6-10?v=New+Era+Pump+Systems+Inc
Average 95 stars, based on 1 article reviews
programmable syringe pump - by Bioz Stars, 2026-08
95/100 stars
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proportional temperature controller  (Scientific Systems Design Inc)
94
Scientific Systems Design Inc proportional temperature controller
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Proportional Temperature Controller, supplied by Scientific Systems Design Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pm16905556-240-41-44?v=Scientific+Systems+Design+Inc
Average 94 stars, based on 1 article reviews
proportional temperature controller - by Bioz Stars, 2026-08
94/100 stars
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cat selb002 igg selb003 igm  (R&D Systems)
90
R&D Systems cat selb002 igg selb003 igm
Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by <t>mRNA</t> signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.
Cat Selb002 Igg Selb003 Igm, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pm27252169-59-32-39?v=R%26D+Systems
Average 90 stars, based on 1 article reviews
cat selb002 igg selb003 igm - by Bioz Stars, 2026-08
90/100 stars
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tension sensor ts module  (Addgene inc)
93
Addgene inc tension sensor ts module
ZO-1 Is under Tensile Stress Regulated by Matrix Stiffness (A) Domain structure of ZO-1. Indicated are the main structural domains and the insertion site of the FRET <t>module.</t> (B and C) The ZO-1 <t>tension</t> <t>sensor</t> (ZO-1-TS) was transiently expressed in Madin-Darby canine kidney (MDCK) cells prior to an analysis of localization by immunofluorescence (B) and immunoblotting (C) with anti-GFP or anti-CTD antibodies. (D and E) FRET analysis by acceptor bleaching and confocal microscopy of full-length ZO-1-TS and the control sensor lacking the CTD. Blebbistatin (10 μM) was added for 20 min prior to imaging. The yellow fluorescent protein (YFP) image taken prior to bleaching reveals the localization of the sensor. The graph shows a quantification of junctional FRET efficiencies of analyzed cells (n for ZO-1-TS, 23; ZO-1-TS-ΔCTD, 17; ZO-1-TS with Blebbistatin, 17; ZO-1-TS-ΔCTD with Blebbistatin, 11; box-plot shows median and interquartile ranges). (F and G) FRET analysis by epifluorescence microscopy of control siRNA-transfected MDCK cells plated on Matrigel-coated glass coverslips or hydrogels of different stiffnesses prior to transfection of the ZO-1 sensor (F). (G) Shows FRET efficiencies at cell-cell contacts of n analyzed cells (n for ZO-1-TS glass, 31; 40 kPa, 31; 1 kPa, 20; ZO-1-TS-ΔCTD glass, 17; 40 kPa, 18; 1 kPa, 18; box-plot shows median and interquartile ranges). Magnification bars, 20 μm. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
Tension Sensor Ts Module, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/pmc07383227-48-0-5?v=Addgene+inc
Average 93 stars, based on 1 article reviews
tension sensor ts module - by Bioz Stars, 2026-08
93/100 stars
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mini protean tetra cell system  (Bio-Rad)
91
Bio-Rad mini protean tetra cell system
ZO-1 Is under Tensile Stress Regulated by Matrix Stiffness (A) Domain structure of ZO-1. Indicated are the main structural domains and the insertion site of the FRET <t>module.</t> (B and C) The ZO-1 <t>tension</t> <t>sensor</t> (ZO-1-TS) was transiently expressed in Madin-Darby canine kidney (MDCK) cells prior to an analysis of localization by immunofluorescence (B) and immunoblotting (C) with anti-GFP or anti-CTD antibodies. (D and E) FRET analysis by acceptor bleaching and confocal microscopy of full-length ZO-1-TS and the control sensor lacking the CTD. Blebbistatin (10 μM) was added for 20 min prior to imaging. The yellow fluorescent protein (YFP) image taken prior to bleaching reveals the localization of the sensor. The graph shows a quantification of junctional FRET efficiencies of analyzed cells (n for ZO-1-TS, 23; ZO-1-TS-ΔCTD, 17; ZO-1-TS with Blebbistatin, 17; ZO-1-TS-ΔCTD with Blebbistatin, 11; box-plot shows median and interquartile ranges). (F and G) FRET analysis by epifluorescence microscopy of control siRNA-transfected MDCK cells plated on Matrigel-coated glass coverslips or hydrogels of different stiffnesses prior to transfection of the ZO-1 sensor (F). (G) Shows FRET efficiencies at cell-cell contacts of n analyzed cells (n for ZO-1-TS glass, 31; 40 kPa, 31; 1 kPa, 20; ZO-1-TS-ΔCTD glass, 17; 40 kPa, 18; 1 kPa, 18; box-plot shows median and interquartile ranges). Magnification bars, 20 μm. See also <xref ref-type=Figure S1 . " width="250" height="auto" />
Mini Protean Tetra Cell System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/module/bio_rxiv__2022__10__17__512532-65-12-16?v=Bio-Rad
Average 91 stars, based on 1 article reviews
mini protean tetra cell system - by Bioz Stars, 2026-08
91/100 stars
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Image Search Results


Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by mRNA signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.

Journal: bioRxiv

Article Title: A Yeast Two-Hybrid Protein Domain Screening Approach for Ebola Virus-Human Protein Interactions Identifies PABPC1 as a Host Factor Required for Replication

doi: 10.64898/2026.03.05.709814

Figure Lengend Snippet: Inhibition of PABPC1 expression reduces EBOV replication. HeLa cells were mock-transfected (reagent only) or transfected with two distinct PABPC1 siRNAs or nontargeting AllStars negative control siRNAs. At 48 h post-transfection, cells were infected with EBOV at an MOI of 0.1. A.) At 16 hpi, samples were inactivated in 10% neutral-buffered formalin and RNAFISH was performed using probes detecting (+) sense NP and VP35 RNA (magenta). Nuclei were visualized by staining with Hoechst (blue). Scale bar = 250 μM. B.) Quantification of NP and VP35 RNA staining in panel A was performed in ImageJ by calculating the area occupied by mRNA signal and normalizing it to the area occupied by Hoechst (nuclei) signal. C.) In a parallel set of samples, total RNA was isolated by lysing the cells with Trizol reagent at 16 hours post infection. NP RNA levels were quantified by one-step RT-qPCR using a (+) sense NP probe to detect EBOV RNA and are normalized to the level of β-Actin RNA. D) Depletion of PABPC1 in siRNA-treated cells. A parallel set of samples were subjected to SDS-PAGE followed by western blotting with PABPC1 and actin antibodies. Molecular weight markers in kDa are shown at left.

Article Snippet: Unidirectional cDNA was prepared from 1 μg of total RNA using the NEBNext® UltraTM Directional RNA Library Prep Kit for Illumina following the “Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490)” through step 1.7 (Manual Version 5.0 5/15) with the following changes: 1) poly(A)+ mRNA was eluted for 5 minutes at 65 °C to prevent fragmentation; 2) after synthesizing second strand cDNA, the cDNA was purified using a Zymo Research DNA Clean & Concentrator-5 column (#D4013); 3) the adaptor primer (/5Phos/GATCGGAAGAGCTTGTTCTACCGAGGGACCC/ideoxyU/ ACTACTGCCTAAC GAACTCCCGCTCTTCCGATC*T; * indicates a phosphorothioate bond; synthesized and PAGE purified by IDT) was a modification of the NEBNext Adaptor for Illumina (E7352A).

Techniques: Inhibition, Expressing, Transfection, Negative Control, Infection, Staining, Isolation, Quantitative RT-PCR, SDS Page, Western Blot, Molecular Weight

ZO-1 Is under Tensile Stress Regulated by Matrix Stiffness (A) Domain structure of ZO-1. Indicated are the main structural domains and the insertion site of the FRET module. (B and C) The ZO-1 tension sensor (ZO-1-TS) was transiently expressed in Madin-Darby canine kidney (MDCK) cells prior to an analysis of localization by immunofluorescence (B) and immunoblotting (C) with anti-GFP or anti-CTD antibodies. (D and E) FRET analysis by acceptor bleaching and confocal microscopy of full-length ZO-1-TS and the control sensor lacking the CTD. Blebbistatin (10 μM) was added for 20 min prior to imaging. The yellow fluorescent protein (YFP) image taken prior to bleaching reveals the localization of the sensor. The graph shows a quantification of junctional FRET efficiencies of analyzed cells (n for ZO-1-TS, 23; ZO-1-TS-ΔCTD, 17; ZO-1-TS with Blebbistatin, 17; ZO-1-TS-ΔCTD with Blebbistatin, 11; box-plot shows median and interquartile ranges). (F and G) FRET analysis by epifluorescence microscopy of control siRNA-transfected MDCK cells plated on Matrigel-coated glass coverslips or hydrogels of different stiffnesses prior to transfection of the ZO-1 sensor (F). (G) Shows FRET efficiencies at cell-cell contacts of n analyzed cells (n for ZO-1-TS glass, 31; 40 kPa, 31; 1 kPa, 20; ZO-1-TS-ΔCTD glass, 17; 40 kPa, 18; 1 kPa, 18; box-plot shows median and interquartile ranges). Magnification bars, 20 μm. See also <xref ref-type=Figure S1 . " width="100%" height="100%">

Journal: Cell Reports

Article Title: Interplay between Extracellular Matrix Stiffness and JAM-A Regulates Mechanical Load on ZO-1 and Tight Junction Assembly

doi: 10.1016/j.celrep.2020.107924

Figure Lengend Snippet: ZO-1 Is under Tensile Stress Regulated by Matrix Stiffness (A) Domain structure of ZO-1. Indicated are the main structural domains and the insertion site of the FRET module. (B and C) The ZO-1 tension sensor (ZO-1-TS) was transiently expressed in Madin-Darby canine kidney (MDCK) cells prior to an analysis of localization by immunofluorescence (B) and immunoblotting (C) with anti-GFP or anti-CTD antibodies. (D and E) FRET analysis by acceptor bleaching and confocal microscopy of full-length ZO-1-TS and the control sensor lacking the CTD. Blebbistatin (10 μM) was added for 20 min prior to imaging. The yellow fluorescent protein (YFP) image taken prior to bleaching reveals the localization of the sensor. The graph shows a quantification of junctional FRET efficiencies of analyzed cells (n for ZO-1-TS, 23; ZO-1-TS-ΔCTD, 17; ZO-1-TS with Blebbistatin, 17; ZO-1-TS-ΔCTD with Blebbistatin, 11; box-plot shows median and interquartile ranges). (F and G) FRET analysis by epifluorescence microscopy of control siRNA-transfected MDCK cells plated on Matrigel-coated glass coverslips or hydrogels of different stiffnesses prior to transfection of the ZO-1 sensor (F). (G) Shows FRET efficiencies at cell-cell contacts of n analyzed cells (n for ZO-1-TS glass, 31; 40 kPa, 31; 1 kPa, 20; ZO-1-TS-ΔCTD glass, 17; 40 kPa, 18; 1 kPa, 18; box-plot shows median and interquartile ranges). Magnification bars, 20 μm. See also Figure S1 .

Article Snippet: Tension sensor (TS) module , Addgene , Cat#26021; RRID:Addgene_26021.

Techniques: Immunofluorescence, Western Blot, Confocal Microscopy, Control, Imaging, Epifluorescence Microscopy, Transfection

Journal: Cell Reports

Article Title: Interplay between Extracellular Matrix Stiffness and JAM-A Regulates Mechanical Load on ZO-1 and Tight Junction Assembly

doi: 10.1016/j.celrep.2020.107924

Figure Lengend Snippet:

Article Snippet: Tension sensor (TS) module , Addgene , Cat#26021; RRID:Addgene_26021.

Techniques: Recombinant