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93
Bio-Rad resolving gel buffer for page
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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Rockland Immunochemicals sds polyacrylamide gel electrophoresis
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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Beyotime sds page gel preparation kit
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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Beyotime beyogeltm plus precast page gel
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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93
Bio-Rad polyacrylamide gel
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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Beyotime dna page gel extraction kit
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
Dna Page Gel Extraction Kit, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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96
Elabscience Biotechnology sds page gel assay kit
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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94
SMOBIO Technology q pagetm bis tris precast gel
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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SMOBIO Technology q pagetm tgn precast gel
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
Q Pagetm Tgn Precast Gel, supplied by SMOBIO Technology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Genesee Scientific gel electrophoresis
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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91
Rockland Immunochemicals 10x sds running buffer
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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93
SMOBIO Technology q page tgn precast gels
a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing <t>gel</t> <t>electrophoresis</t> with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).
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Image Search Results


a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing gel electrophoresis with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).

Journal: Nature

Article Title: SIGLEC12 mediates plasma membrane rupture during necroptotic cell death

doi: 10.1038/s41586-025-09741-1

Figure Lengend Snippet: a – d , HT - 29 cells with the indicated knockouts ( a ) or knockdowns ( b – d ) were treated with TSE (30 ng ml −1 hTNF, 0.2 µM SM-164, 5 µM emricasan) or TRAIL + SE (50 ng ml −1 , 0.2 µM SM-164, 5 µM emricasan) for 8 h, and cell death or cell viability was quantified using the indicated assays ( P = 1.39 × 10 −13 ( a ), P < 1 × 10 −15 ( b ), P < 1 × 10 −15 ( c ) and P = 6.82 × 10 −12 , P = 1.23 × 10 −11 ( d )). Cell lysates were analysed using reducing or non-reducing gel electrophoresis with the indicated antibodies. e , HT-29 cells with indicated stable short hairpin RNA (shRNA)-mediated knockdowns were treated with TSE for the indicated times, and cell death was assessed using CellToxGreen and time-lapse fluorescence confocal microscopy. f , As in e , except cell morphology was visualized by scanning electron microscopy. g , As in e , except the release of HMGB1 was analysed by immunoblotting. Data are plotted as the mean ± s.d., representative of n = 4 ( c ), n = 9 ( a , d ) or n = 12 ( b ); three independent experiments. Statistical analyses were performed using two-tailed t -tests and two-way analyses of variance; **** P < 0.0001. Data are representative of three independent experiments ( d , middle and right, g ). Scale bars, 25 μm ( e ), 10 μm ( f ).

Article Snippet: The following reagents were used in this study: SM-164 (S7089), Nec-1s (S8641), GSK′872 (S8465), NSA (S8251), emricasan (S7775) and erastin (S7242) from SelleckChem; etoposide (E1383), α-ketoglutarate (349631), lipopolysaccharide (L4391), luminol (A8511), p -coumaric acid (C9008) and anti-FLAG M2 agarose beads (M8823) from Sigma-Aldrich; nigericin (11437) from the Cayman Chemical Company; Lipofectamine 3000 (L3000015), SuperSignal West Atto Ultimate Sensitivity Substrate (A38556) and Prolong Diamond Antifade Mountant with DAPI ( P36966 ) from Thermo Fisher Scientific; polyethylenimine (PEI; 24765-100) from Kyfora Bio; polybrene (TR-1003) from EMD Millipore; and 10X Tris/Glycine/SDS Electrophoresis Buffer (1610772), Tween 20 (1610781), Stacking Gel Buffer for PAGE (1610799), Resolving Gel Buffer for PAGE (1610798), Precision Plus Protein Dual Color Standards (1610394) and nitrocellulose membrane (1620115) from Bio-Rad.

Techniques: Nucleic Acid Electrophoresis, shRNA, Fluorescence, Confocal Microscopy, Electron Microscopy, Western Blot, Two Tailed Test