nec 1 Search Results


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MedChemExpress nec
Nec, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TargetMol necrostatin 1
Necrostatin 1, supplied by TargetMol, 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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MedChemExpress necrostatin 1
Necrostatin 1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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MedChemExpress ogd r necrostatin 1
Ogd R Necrostatin 1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ProSci Incorporated nec
Publications concerning necroptosis in liver injury from acetaminophen toxicity
Nec, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93
OriGene pcsk1 myc ddk
a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of <t>Pro-Pcsk1</t> and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.
Pcsk1 Myc Ddk, supplied by OriGene, 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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Fluorochem Ltd necroptosis inhibitors nec-1
a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of <t>Pro-Pcsk1</t> and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.
Necroptosis Inhibitors Nec 1, supplied by Fluorochem Ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Beyotime necrostatin-1
a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of <t>Pro-Pcsk1</t> and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.
Necrostatin 1, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Merck KGaA rip1 inhibitor 7-cl-o-nec-1
a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of <t>Pro-Pcsk1</t> and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.
Rip1 Inhibitor 7 Cl O Nec 1, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Enzo Biochem nec 1
PEG‐tAHP‐DRI inhibited TC‐induced HUVECs necroptosis in vitro. A) TCs transmigration over the HUVEC monolayer after a treatment with <t>zVAD,</t> <t>Nec‐1,</t> anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. B) HUVECs necroptosis after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. C) Quantification of transmigration TCs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. D) Quantification of necroptosis HUVECs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. Data are means ± SD ( n = 3), and analyzed with GraphPad Prism 8.0. * p < 0.05, ** p < 0.01, and *** p < 0.005 calculated by unpaired two‐tailed Student's t ‐test. NS indicates p > 0.05.
Nec 1, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Oncoscience ag necroptosis-specific inhibitor, necrostatin-1 (nec-1)
PEG‐tAHP‐DRI inhibited TC‐induced HUVECs necroptosis in vitro. A) TCs transmigration over the HUVEC monolayer after a treatment with <t>zVAD,</t> <t>Nec‐1,</t> anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. B) HUVECs necroptosis after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. C) Quantification of transmigration TCs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. D) Quantification of necroptosis HUVECs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. Data are means ± SD ( n = 3), and analyzed with GraphPad Prism 8.0. * p < 0.05, ** p < 0.01, and *** p < 0.005 calculated by unpaired two‐tailed Student's t ‐test. NS indicates p > 0.05.
Necroptosis Specific Inhibitor, Necrostatin 1 (Nec 1), supplied by Oncoscience ag, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Merck & Co nec-1 s (7-cl − o- nec - 1
PEG‐tAHP‐DRI inhibited TC‐induced HUVECs necroptosis in vitro. A) TCs transmigration over the HUVEC monolayer after a treatment with <t>zVAD,</t> <t>Nec‐1,</t> anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. B) HUVECs necroptosis after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. C) Quantification of transmigration TCs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. D) Quantification of necroptosis HUVECs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. Data are means ± SD ( n = 3), and analyzed with GraphPad Prism 8.0. * p < 0.05, ** p < 0.01, and *** p < 0.005 calculated by unpaired two‐tailed Student's t ‐test. NS indicates p > 0.05.
Nec 1 S (7 Cl − O Nec 1, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Publications concerning necroptosis in liver injury from acetaminophen toxicity

Journal: Cell Death Discovery

Article Title: Questions and controversies: the role of necroptosis in liver disease

doi: 10.1038/cddiscovery.2016.89

Figure Lengend Snippet: Publications concerning necroptosis in liver injury from acetaminophen toxicity

Article Snippet: Ramachandran et al . , RIPK3 morpholino protected against APAP 200 mg/kg at 6 h Nec-1 protected in vitro RIPK3 induction suggested with APAP RIPK3−/− protected at 6hr No protection at 24 h in RIPK3−/− , Polyclonal antibody Prosci Mismatched strain as controls and not littermate controls Used Morpholino to knockdown but was not detecting RIPK3 at baseline suggesting RIPK3 induction prevented with morpholino pre-treatment RIPK3 present at baseline inconsistently on blots Nec-1 s (more specific inhibitor) not used Only early time point protection No effect on APAP metabolism (GSH unchanged between tx and con).

Techniques: Molecular Weight, In Vitro, Knockdown, Immunohistochemistry, Injection, Modification, In Vivo, Binding Assay, Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Staining, Membrane

a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Verification of loss of CerS2 in CerS2 ΔIns1E cells by immunoblot. Left: representative immunoblot. Right: quantification of Cers2 signals ( n = 8 independent experiments). b , Representative immunostaining (left) and quantification (right) of ER marker PDI in control and CerS2 ΔIns1E cells ( n = 36 control versus 28 CerS2 ΔIns1E well sites from one experiment). Scale bar, 10 μm. c , Quantification of insulin content in control and CerS2 ΔIns1E cells at low (2 mM) and high (25 mM) glucose levels ( n = 5 independent experiments). d , Experimental design and results for proteome analyses in control and CerS2 ΔIns1E cells. e , Volcano plot showing log 2 fold change of proteins between CerS2 ΔIns1E and control cells plotted against the −log 10 P value of a two-sided paired Student’s t -test. BH-FDR <0.05 and fold change >1.5 was used as significance cut-offs ( n = 3 control versus 3 CerS2 ΔIns1E samples collected in three independent experiments). f , g , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of control and CerS2 ΔBKO mice. f , Representative immunoblot. Each lane represents islets of one individual mouse. g , Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six CerS2 ΔBKO mice). h , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and db/db.BKS mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six db/db.BKS mice). Representative immunoblot is shown in Extended Data Fig. . i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in islets of 12-week-old control and ob/ob.B6 mice. Quantification of Pro-Pcsk1 (left) and Pcsk1 (middle) protein levels and ratio of Pcsk1/Pro-Pcsk1 (right; n = islets of six control versus six ob/ob.B6 mice). Representative immunoblot is shown in Extended Data Fig. . Statistical analysis was performed using a two-sided Student’s t -test ( a , b , g , h and i ) and two-way ANOVA with Sidak’s multiple comparisons test ( c ). P values are stated in each figure. Bar graphs represent mean ( c ) or mean ± s.e.m. ( a , b , g , h and i ). Connecting lines indicate both samples are from one experiment. Data points in b represent individual well sites. Data points in a and c represent independent experiments. Data points in g – i represent islets from individual mice. Stain-free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: Western Blot, Immunostaining, Marker, Staining

a , Quantification of Pcsk1 levels in control and CerS2 ΔIns1E cells by CrispR-verified antibody (Supplementary Fig. , Cell Signaling #11914, discontinued). Left, representative immunoblot. Right, quantification of Pcsk1 signals ( n = 5 independent experiments). b , Quantification of Pcsk1 protein levels in control and CerS2 BKO islets by CrispR-verified antibody (Supplementary Fig. , Cell Signaling #11914, discontinued). Left, representative immunoblot. Right, quantification of Pcsk1 signals ( n = 8 independent experiments). c , Quantification of Pcsk1 mRNA levels in islets from male control and CerS2 BKO mice by qPCR ( n = 5 independent experiments). d , Representative immunoblots to Fig. . e , Representative immunoblots to Fig. . f, g , Quantification of mRNA levels of Pcsk1 and various CerS in islets of 12 week old control and db/db.BKS mice (f, n = 4 control and 4 db.db/BKS islet samples) and islets of 12 week old control and ob/ob.B6 mice (g, n = 4 control and 6 ob/ob.B6 islets samples). Statistical analysis was performed using a paired two-sided Student’s t -test (a, b), two-sided Student’s t -test (c) and two-sided multiple t -tests with Holm-Sidak correction (f, g). P -values are stated in each figure. Bar graphs represent means (a, b) or means + s.e.m. (c, f, g). Connecting lines indicate both samples are from one experiment. Data points represent independent experiments (a), islets from individual mice (c) or individual islet samples (f, g). Stain-Free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Quantification of Pcsk1 levels in control and CerS2 ΔIns1E cells by CrispR-verified antibody (Supplementary Fig. , Cell Signaling #11914, discontinued). Left, representative immunoblot. Right, quantification of Pcsk1 signals ( n = 5 independent experiments). b , Quantification of Pcsk1 protein levels in control and CerS2 BKO islets by CrispR-verified antibody (Supplementary Fig. , Cell Signaling #11914, discontinued). Left, representative immunoblot. Right, quantification of Pcsk1 signals ( n = 8 independent experiments). c , Quantification of Pcsk1 mRNA levels in islets from male control and CerS2 BKO mice by qPCR ( n = 5 independent experiments). d , Representative immunoblots to Fig. . e , Representative immunoblots to Fig. . f, g , Quantification of mRNA levels of Pcsk1 and various CerS in islets of 12 week old control and db/db.BKS mice (f, n = 4 control and 4 db.db/BKS islet samples) and islets of 12 week old control and ob/ob.B6 mice (g, n = 4 control and 6 ob/ob.B6 islets samples). Statistical analysis was performed using a paired two-sided Student’s t -test (a, b), two-sided Student’s t -test (c) and two-sided multiple t -tests with Holm-Sidak correction (f, g). P -values are stated in each figure. Bar graphs represent means (a, b) or means + s.e.m. (c, f, g). Connecting lines indicate both samples are from one experiment. Data points represent independent experiments (a), islets from individual mice (c) or individual islet samples (f, g). Stain-Free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: CRISPR, Western Blot, Staining

a , Immunoblot detection of human CerS6 (hCerS6) and Tmed2 protein levels in INS1E cells after overexpression of hCerS6 via adenovirus for 48 h (representative immunoblot). b , Ins1E cell counts after 48 hours of infection with a control adenovirus or hCerS6-expressing adenovirus ( n = 3 independent experiments). A reduction of cell counts is in line with the ability of CerS6 overexpression to induce apoptosis in several cell types. c , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels (Cell Signaling #18030) in Ins1E cells infected with a control adenovirus or hCerS6-expressing adenovirus for 48 h (representative immunoblot). d-g , Quantification of Tmed2 (d), Pcsk1 (e), Pro-Pcsk1 signals (f) and Pcsk1/ Pro-Pcsk1 ratio (g) from 4 independent experiments. Statistical analysis was performed using two-way ANOVA with Sidak’s multiple comparisons test (b, d-g). Data points in (b, d-g) represent independent experiments. Bar graphs in (b, d-g) represent means + s.e.m. Stain-Free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Immunoblot detection of human CerS6 (hCerS6) and Tmed2 protein levels in INS1E cells after overexpression of hCerS6 via adenovirus for 48 h (representative immunoblot). b , Ins1E cell counts after 48 hours of infection with a control adenovirus or hCerS6-expressing adenovirus ( n = 3 independent experiments). A reduction of cell counts is in line with the ability of CerS6 overexpression to induce apoptosis in several cell types. c , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels (Cell Signaling #18030) in Ins1E cells infected with a control adenovirus or hCerS6-expressing adenovirus for 48 h (representative immunoblot). d-g , Quantification of Tmed2 (d), Pcsk1 (e), Pro-Pcsk1 signals (f) and Pcsk1/ Pro-Pcsk1 ratio (g) from 4 independent experiments. Statistical analysis was performed using two-way ANOVA with Sidak’s multiple comparisons test (b, d-g). Data points in (b, d-g) represent independent experiments. Bar graphs in (b, d-g) represent means + s.e.m. Stain-Free signal was used for normalization of all immunoblots. Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: Western Blot, Over Expression, Infection, Expressing, Staining

a , Experimental setup for identification of SBPs in a SILAC-based approach. pacSph treatment of Sgpl1 ΔIns1E and CerS2:Sgpl1 ΔIns1E cells differentially labelled with stable isotopes allows crosslinking of SL-protein complexes by UV irradiation (with omission of UV irradiation as a control condition), followed by cell lysis and conjugation of biotin to the SL-protein complexes. After Streptavidin-based pull-down, SBPs can be identified and quantified in the same MS run by the differing peptide mass due to SILAC isotope labelling. b , Volcano plot showing log 2 fold change of proteins pulled down from pacSph-treated Sgpl1 ΔIns1E (+UV) versus Sgpl1 ΔIns1E (−UV) cells plotted against the −log 10 P values of a one-sample two-sided t -test against 0. Proteins with log 2 fold change >1 and a BH-FDR <0.05 are regarded as SBPs ( n = 4 independent experiments). c , Volcano plot showing log 2 fold change of SBPs identified in b (Supplementary Fig. ) and pulled down from pacSph-treated CerS2:Sgpl1 ΔIns1E (+UV) versus Sgpl1 ΔIns1E (+UV) cells plotted against the −log 10 P values of a two-sample two-sided equal variance t -test ( n = 4 independent experiments). SBPs with a fold change >1.5 and a BH-FDR <0.05 were regarded as Cers2-dependent SBPs. Fold enrichment was 6.55 and FDR-corrected P value was 3.12 −10 for GO term ‘endoplasmic reticulum’. d , pacSph pull-down of endogenous Tmed2 in Sgpl ΔIns1E and Sgpl1:CerS2 ΔIns1E cells ( n = 4 independent experiments); exemplary immunoblot (right) and quantification (left). Eluate intensities were normalized to respective input intensities. e , Relative mRNA expression of Tmed1, Tmed2 and Pcsk1 in murine pseudoislets transfected with control siRNA or siRNA against Tmed1, Tmed2 or both. n = 4 independent experiments. f – i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in pseudoislets transfected with siRNA as described in Extended Data Fig. ; n = 3 independent experiments. f , Representative immunoblot. g , Quantification of Pcsk1. h , Quantification of Pro-Pcsk1. i , Ratio of Pcsk1 to Pro-Pcsk1. j – l , Insulin content ( j ), proinsulin content ( k ) and ratio of insulin to proinsulin ( l ) in pseudoislets transfected with siRNA as described in Extended Data Fig. determined via ELISA ( n = 8 independent experiments). Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparisons test ( d ) and repeated measures one-way ANOVA with Tukey’s multiple comparisons test ( e and g – l ). In e , ANOVA was performed for each mRNA target individually. P values are stated in each figure. Data points in d , e and g – l represent individual experiments. Bar graphs represent mean ± s.e.m. For one experiment in j – l , the mean of five replicates, consisting of nine pseudoislets, respectively, was plotted per condition. Stain-free signal was used for normalization of immunoblots in g – i . Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Experimental setup for identification of SBPs in a SILAC-based approach. pacSph treatment of Sgpl1 ΔIns1E and CerS2:Sgpl1 ΔIns1E cells differentially labelled with stable isotopes allows crosslinking of SL-protein complexes by UV irradiation (with omission of UV irradiation as a control condition), followed by cell lysis and conjugation of biotin to the SL-protein complexes. After Streptavidin-based pull-down, SBPs can be identified and quantified in the same MS run by the differing peptide mass due to SILAC isotope labelling. b , Volcano plot showing log 2 fold change of proteins pulled down from pacSph-treated Sgpl1 ΔIns1E (+UV) versus Sgpl1 ΔIns1E (−UV) cells plotted against the −log 10 P values of a one-sample two-sided t -test against 0. Proteins with log 2 fold change >1 and a BH-FDR <0.05 are regarded as SBPs ( n = 4 independent experiments). c , Volcano plot showing log 2 fold change of SBPs identified in b (Supplementary Fig. ) and pulled down from pacSph-treated CerS2:Sgpl1 ΔIns1E (+UV) versus Sgpl1 ΔIns1E (+UV) cells plotted against the −log 10 P values of a two-sample two-sided equal variance t -test ( n = 4 independent experiments). SBPs with a fold change >1.5 and a BH-FDR <0.05 were regarded as Cers2-dependent SBPs. Fold enrichment was 6.55 and FDR-corrected P value was 3.12 −10 for GO term ‘endoplasmic reticulum’. d , pacSph pull-down of endogenous Tmed2 in Sgpl ΔIns1E and Sgpl1:CerS2 ΔIns1E cells ( n = 4 independent experiments); exemplary immunoblot (right) and quantification (left). Eluate intensities were normalized to respective input intensities. e , Relative mRNA expression of Tmed1, Tmed2 and Pcsk1 in murine pseudoislets transfected with control siRNA or siRNA against Tmed1, Tmed2 or both. n = 4 independent experiments. f – i , Immunoblot detection of Pro-Pcsk1 and Pcsk1 protein levels in pseudoislets transfected with siRNA as described in Extended Data Fig. ; n = 3 independent experiments. f , Representative immunoblot. g , Quantification of Pcsk1. h , Quantification of Pro-Pcsk1. i , Ratio of Pcsk1 to Pro-Pcsk1. j – l , Insulin content ( j ), proinsulin content ( k ) and ratio of insulin to proinsulin ( l ) in pseudoislets transfected with siRNA as described in Extended Data Fig. determined via ELISA ( n = 8 independent experiments). Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparisons test ( d ) and repeated measures one-way ANOVA with Tukey’s multiple comparisons test ( e and g – l ). In e , ANOVA was performed for each mRNA target individually. P values are stated in each figure. Data points in d , e and g – l represent individual experiments. Bar graphs represent mean ± s.e.m. For one experiment in j – l , the mean of five replicates, consisting of nine pseudoislets, respectively, was plotted per condition. Stain-free signal was used for normalization of immunoblots in g – i . Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: Irradiation, Lysis, Conjugation Assay, Western Blot, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Staining

a , Top 20 SBPs identified in Fig. according to p -values and a log 2 fold change > 3. b , Verification of Bst2 and Fxyd6 as SBPs by overexpression of DDK-tagged variants in Sgpl1 ΔIns1E cells followed by pacSph-pulldown; representative immunoblots (left) and quantification (right). Eluate band intensities were normalized to input bands and +UV samples were set to 1 ( n = 4 independent experiments). c , Verification of Tmed1 as SBP as described in (b); representative immunoblot (left) and quantification (right). n = 3 independent experiments. d , Immunoblot detection of Tmed2 protein levels in Sgpl1 ΔIns1E and Cers2:Sgpl1 ΔIns1E cells. Representative immunoblot showing 3 replicates per genotype (left) and quantification (right). n = 3 independent experiments with 3 replicates per genotype, respectively. e , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 Cers2 ΔBKO mice. Representative immunoblot (left) and quantification (right). f , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 ob/ob.B6 mice at week 12. Representative immunoblot (left) and quantification (right). g , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 db/db.BKS mice at week 12. Representative immunoblot (left) and quantification (right). Statistical analysis was performed using a one sample t -test against 1 (b, c) or Student’s t -test (d-g). P -values are stated in each figure. Bar graphs represent means + s.e.m. Data points in (b, c and d) represent individual experiments. Data points in (e-g) represent islets from individual mice. Stain-Free signal was used for normalization of all immunoblots, except for (b) and (c). Stain-Free images of (e), (f) and (g) were reproduced from Fig. , Extended Data Fig. , as the same PVDF membranes were used for detection of Pro-Pcsk1, Pcsk1 and Tmed2, respectively. Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Top 20 SBPs identified in Fig. according to p -values and a log 2 fold change > 3. b , Verification of Bst2 and Fxyd6 as SBPs by overexpression of DDK-tagged variants in Sgpl1 ΔIns1E cells followed by pacSph-pulldown; representative immunoblots (left) and quantification (right). Eluate band intensities were normalized to input bands and +UV samples were set to 1 ( n = 4 independent experiments). c , Verification of Tmed1 as SBP as described in (b); representative immunoblot (left) and quantification (right). n = 3 independent experiments. d , Immunoblot detection of Tmed2 protein levels in Sgpl1 ΔIns1E and Cers2:Sgpl1 ΔIns1E cells. Representative immunoblot showing 3 replicates per genotype (left) and quantification (right). n = 3 independent experiments with 3 replicates per genotype, respectively. e , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 Cers2 ΔBKO mice. Representative immunoblot (left) and quantification (right). f , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 ob/ob.B6 mice at week 12. Representative immunoblot (left) and quantification (right). g , Immunoblot detection of Tmed2 protein levels in islets of 6 control and 6 db/db.BKS mice at week 12. Representative immunoblot (left) and quantification (right). Statistical analysis was performed using a one sample t -test against 1 (b, c) or Student’s t -test (d-g). P -values are stated in each figure. Bar graphs represent means + s.e.m. Data points in (b, c and d) represent individual experiments. Data points in (e-g) represent islets from individual mice. Stain-Free signal was used for normalization of all immunoblots, except for (b) and (c). Stain-Free images of (e), (f) and (g) were reproduced from Fig. , Extended Data Fig. , as the same PVDF membranes were used for detection of Pro-Pcsk1, Pcsk1 and Tmed2, respectively. Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: Over Expression, Western Blot, Staining

a , Double cut CrispR/Cas9 knockout strategy for Tmed2 in Ins1E cells. b, c , Relative mRNA expression of various Tmed family members (b) and beta cell identity markers (c) in wildtype Ins1E, control and Tmed2 ΔIns1E cells. Wildtype Ins1E samples were set to 1 (dotted line). n = 3 independent experiments. Note that potentially as a sign of attempted compensation, Pcsk1 mRNA levels are increased in Tmed2-deficient Ins1E cells. d , Representative immunoblot analysis of Tmed2 protein expression in 3 control vs. 3 Tmed2 ΔIns1E replicate lysates. e , Insulin content in control and Tmed2 ΔIns1E cells determined via ELISA. n = 3 independent experiments with 3 replicates per genotype, respectively. Statistical analysis was performed using multiple two-sided t -tests with Holm-Sidak correction (b and c) and a two-sided Student’s t -test (e). Data points represent independent experiments. Bar graphs represent means + s.e.m. The control and Tmed2 ΔIns1E cells are pools of individual monoclonal cell lines, respectively (8 monoclonal control cell lines and 3 monoclonal Tmed2 ΔIns1E cell lines were used for pooling). Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Double cut CrispR/Cas9 knockout strategy for Tmed2 in Ins1E cells. b, c , Relative mRNA expression of various Tmed family members (b) and beta cell identity markers (c) in wildtype Ins1E, control and Tmed2 ΔIns1E cells. Wildtype Ins1E samples were set to 1 (dotted line). n = 3 independent experiments. Note that potentially as a sign of attempted compensation, Pcsk1 mRNA levels are increased in Tmed2-deficient Ins1E cells. d , Representative immunoblot analysis of Tmed2 protein expression in 3 control vs. 3 Tmed2 ΔIns1E replicate lysates. e , Insulin content in control and Tmed2 ΔIns1E cells determined via ELISA. n = 3 independent experiments with 3 replicates per genotype, respectively. Statistical analysis was performed using multiple two-sided t -tests with Holm-Sidak correction (b and c) and a two-sided Student’s t -test (e). Data points represent independent experiments. Bar graphs represent means + s.e.m. The control and Tmed2 ΔIns1E cells are pools of individual monoclonal cell lines, respectively (8 monoclonal control cell lines and 3 monoclonal Tmed2 ΔIns1E cell lines were used for pooling). Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: CRISPR, Knock-Out, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

a , Co-immunoprecipitation (Co-IP) of co-overexpressed Tmed2-V5 and Pro-Pcsk1/Pcsk1-DDK in Ins1E cells. Representative immunoblot (left) and quantification of three replicate experiments (right). As Ctrl-plasmid, the promotorless pNL1.3 from Promega (N1021) was used. b , Representative confocal images for co-localization of overexpressed Tmed2-V5 and Pro-/Pcsk1-DDK in Ins1E cells. Green, SytoxGreen as nucleus marker; red, Pro-/Pcsk1-DDK; blue, Tmed2-V5. Scale bar, 5 µm. c, d , Quantification of overlap of Pro-/Pcsk1-DDK with Tmed2-V5 (c) and Tmed2-V5 with Pro-/Pcsk1-DDK (d) in control and CerS2 ΔIns1E cells. n = 2 independent experiments; only one experiment shown. e-j , Overlap of ER-marker PDI and Golgi-Marker TGN46 with Tmed2-V5, Pro-/Pcsk1-DDK (allowing detection of both Pro-Pcsk1 as well as mature Pcsk1) and Pro-Pcsk1 (only allowing detection of the immature Pro-Pcsk1 protein) after overexpression in control and CerS2 ΔIns1E cells. n = 3 independent experiments. Statistical analysis was performed using a paired two-sided Student’s t -test (a) and unpaired two-sided Students t -tests (c-j). Data points represent replicate experiments (a) and individually quantified cells (c-d) or well sites (e-j). Bar graphs represent means + s.e.m. Source numerical data and unprocessed blots are available in source data.

Journal: Nature Cell Biology

Article Title: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis

doi: 10.1038/s41556-022-01027-2

Figure Lengend Snippet: a , Co-immunoprecipitation (Co-IP) of co-overexpressed Tmed2-V5 and Pro-Pcsk1/Pcsk1-DDK in Ins1E cells. Representative immunoblot (left) and quantification of three replicate experiments (right). As Ctrl-plasmid, the promotorless pNL1.3 from Promega (N1021) was used. b , Representative confocal images for co-localization of overexpressed Tmed2-V5 and Pro-/Pcsk1-DDK in Ins1E cells. Green, SytoxGreen as nucleus marker; red, Pro-/Pcsk1-DDK; blue, Tmed2-V5. Scale bar, 5 µm. c, d , Quantification of overlap of Pro-/Pcsk1-DDK with Tmed2-V5 (c) and Tmed2-V5 with Pro-/Pcsk1-DDK (d) in control and CerS2 ΔIns1E cells. n = 2 independent experiments; only one experiment shown. e-j , Overlap of ER-marker PDI and Golgi-Marker TGN46 with Tmed2-V5, Pro-/Pcsk1-DDK (allowing detection of both Pro-Pcsk1 as well as mature Pcsk1) and Pro-Pcsk1 (only allowing detection of the immature Pro-Pcsk1 protein) after overexpression in control and CerS2 ΔIns1E cells. n = 3 independent experiments. Statistical analysis was performed using a paired two-sided Student’s t -test (a) and unpaired two-sided Students t -tests (c-j). Data points represent replicate experiments (a) and individually quantified cells (c-d) or well sites (e-j). Bar graphs represent means + s.e.m. Source numerical data and unprocessed blots are available in source data.

Article Snippet: To determine overlap of Tmed2 and Pcsk1 with ER and Golgi, we used the following plasmids: Tmed2-Myc-DDK (Origene, #RC206849), Pcsk1-Myc-DDK (DDK-tag will be cleaved off during maturation into enzymatically active Pcsk1; Origene, #MR225451) and the Pro-/Pcsk1-DDK plasmid described above.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Plasmid Preparation, Marker, Over Expression

PEG‐tAHP‐DRI inhibited TC‐induced HUVECs necroptosis in vitro. A) TCs transmigration over the HUVEC monolayer after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. B) HUVECs necroptosis after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. C) Quantification of transmigration TCs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. D) Quantification of necroptosis HUVECs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. Data are means ± SD ( n = 3), and analyzed with GraphPad Prism 8.0. * p < 0.05, ** p < 0.01, and *** p < 0.005 calculated by unpaired two‐tailed Student's t ‐test. NS indicates p > 0.05.

Journal: Advanced Science

Article Title: Multi‐Arm PEG/Peptidomimetic Conjugate Inhibitors of DR6/APP Interaction Block Hematogenous Tumor Cell Extravasation

doi: 10.1002/advs.202003558

Figure Lengend Snippet: PEG‐tAHP‐DRI inhibited TC‐induced HUVECs necroptosis in vitro. A) TCs transmigration over the HUVEC monolayer after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. B) HUVECs necroptosis after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. C) Quantification of transmigration TCs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. D) Quantification of necroptosis HUVECs after a treatment with zVAD, Nec‐1, anti‐DR6 antibody, AHP‐DRI‐12, or PEG‐tAHP‐DRI. Data are means ± SD ( n = 3), and analyzed with GraphPad Prism 8.0. * p < 0.05, ** p < 0.01, and *** p < 0.005 calculated by unpaired two‐tailed Student's t ‐test. NS indicates p > 0.05.

Article Snippet: Nec‐1 and z‐VAD were purchased from Enzo Biochem (New York, USA).

Techniques: In Vitro, Transmigration Assay, Two Tailed Test