multiple gaussian functions originpro 2020 Search Results


multiple gaussian functions originpro 2020  (OriginLab corp)
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resource source identifier antibodies nf45 bethyl  (Bethyl)
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Figure 2. The <t>NF45-NF90</t> Complex Is Required for Mitotic Progression and Cytokinesis (A) Proliferation assay using WST-1 of HeLa cells with stable depletion of NF45 or NF90 over a 72-h time course. Data are expressed as the mean fold-change in optical density compared to the 0-h time point (A.U., arbitrary units). (B) Fluorescence-activated cell sorting (FACS) analysis of DNA content of HeLa cells stably expressing shNT, shNF45, or shNF90. Data analysis and quantifi- cation of cells at different phase of the cell cycle were done using Flowjo. (C) Quantification of binucleated cells by immunofluorescence in response to NF45 or NF90 depletion. A minimum of 1,500 cells per condition was quantified. ***p < 0.001 (two-tailed t test).
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xtalab synergy diffractometer with a dualflex hypix detector  (Rigaku Corporation)
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Figure 2. The <t>NF45-NF90</t> Complex Is Required for Mitotic Progression and Cytokinesis (A) Proliferation assay using WST-1 of HeLa cells with stable depletion of NF45 or NF90 over a 72-h time course. Data are expressed as the mean fold-change in optical density compared to the 0-h time point (A.U., arbitrary units). (B) Fluorescence-activated cell sorting (FACS) analysis of DNA content of HeLa cells stably expressing shNT, shNF45, or shNF90. Data analysis and quantifi- cation of cells at different phase of the cell cycle were done using Flowjo. (C) Quantification of binucleated cells by immunofluorescence in response to NF45 or NF90 depletion. A minimum of 1,500 cells per condition was quantified. ***p < 0.001 (two-tailed t test).
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Image Search Results


Figure 2. The NF45-NF90 Complex Is Required for Mitotic Progression and Cytokinesis (A) Proliferation assay using WST-1 of HeLa cells with stable depletion of NF45 or NF90 over a 72-h time course. Data are expressed as the mean fold-change in optical density compared to the 0-h time point (A.U., arbitrary units). (B) Fluorescence-activated cell sorting (FACS) analysis of DNA content of HeLa cells stably expressing shNT, shNF45, or shNF90. Data analysis and quantifi- cation of cells at different phase of the cell cycle were done using Flowjo. (C) Quantification of binucleated cells by immunofluorescence in response to NF45 or NF90 depletion. A minimum of 1,500 cells per condition was quantified. ***p < 0.001 (two-tailed t test).

Journal: Cell reports

Article Title: NF45 and NF90 Regulate Mitotic Gene Expression by Competing with Staufen-Mediated mRNA Decay.

doi: 10.1016/j.celrep.2020.107660

Figure Lengend Snippet: Figure 2. The NF45-NF90 Complex Is Required for Mitotic Progression and Cytokinesis (A) Proliferation assay using WST-1 of HeLa cells with stable depletion of NF45 or NF90 over a 72-h time course. Data are expressed as the mean fold-change in optical density compared to the 0-h time point (A.U., arbitrary units). (B) Fluorescence-activated cell sorting (FACS) analysis of DNA content of HeLa cells stably expressing shNT, shNF45, or shNF90. Data analysis and quantifi- cation of cells at different phase of the cell cycle were done using Flowjo. (C) Quantification of binucleated cells by immunofluorescence in response to NF45 or NF90 depletion. A minimum of 1,500 cells per condition was quantified. ***p < 0.001 (two-tailed t test).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NF45 Bethyl Cat# A303-147A, RRID:AB_10895652 NF90 Bethyl Cat# A303-651A, RRID:AB_11204576 NUF2 Abcam Cat# ab122962, RRID:AB_10902068 CDK1 Santa Cruz Cat# sc-54, RRID:AB_627224 CCNA2 Santa Cruz Cat# sc-53229, RRID:AB_782331 Survivin Novus Biological Cat# NB500-201, RRID:AB_10001517 Tubulin Sigma-Aldrich Cat# T5168, RRID:AB_477579 Actin Sigma-Aldrich Cat# A5441, RRID:AB_476744 Myc Millipore Cat# 05-419, RRID:AB_309725 UPF1 MBL International Cat# RN108PW, RRID:AB_2616172 p-H3 Serine 10 Cell Signaling Cat# 3377, RRID:AB_1549592 HA tag Sigma-Aldrich Cat# H6533, RRID:AB_439705 Mouse-HRP Millipore Cat# 71045-3, RRID:AB_10808067 Rabbit-HRP Millipore Cat# AP304P, RRID:AB_11212320 Tubulin Alexa Fluor 488 Thermo Fisher Scientific Cat# 322588, RRID:AB_2532182 Bacterial and Virus Strains DH5a Thermo Fisher Scientific Cat# 18265017 Experimental Models: Cell Lines HeLa ATCC Cat# CCL-2, RRID:CVCL_0030 HEK293 ATCC Cat# CRL-1573, RRID:CVCL_0045 HEK293T ATCC Cat# CRL-3216, RRID:CVCL_0063 MDA-MB-231 ATCC Cat# CRL-12532, RRID:CVCL_0062 BT-20 ATCC Cat# CRL-7912, RRID:CVCL_0178 Chemicals, Peptides, and Recombinant Proteins Nocodazole Sigma-Aldrich Cat# M1404 Thymidine Sigma-Aldrich Cat# T9250 Puromycin Sigma-Aldrich Cat# P8833 DAPI VECTASHIELD Vector laboratories Cat# H-1200 Coelenterazine-400a Biotium Cat# 10125 Polybrene Sigma-Aldrich Cat# S2667 WST-1 Sigma-Aldrich Cat# 05 015 944 001 Propidium Iodide Sigma-Aldrich Cat# P4170 RNase A Sigma-Aldrich Cat# R6513 Cycloheximide Sigma-Aldrich Biotin Thermo Fisher Scientific Cat# B1595 Tetracycline Sigma-Aldrich Cat# T7660 Hygromycin Sigma-Aldrich Cat# H3274 Protein A Sepharose GE healthcare Cat# 17-0780-01 Critical Commercial Assays RNeasy minikit Quiagen Cat# 74106 Secrete-Pair Gaussia Luciferase Assay Kit Genecopeia Cat# LF032 Luciferase Assay System Promega Cat# E1500 (Continued on next page) Cell Reports 31, 107660, May 19, 2020 e1

Techniques: Proliferation Assay, Fluorescence, FACS, Stable Transfection, Expressing, Two Tailed Test

Figure 3. The NF45-NF90 Complex Regulates the Expression of a Cluster of Mitotic Genes (A) Transcriptome analysis of HepG2 cells depleted for NF45/ILF2. Red dots indicate genes that are significantly depleted by more than 2-fold change compared to control. (B) Classification of mRNAs significantly decreased in HepG2 cells depleted for NF45, based on Reactome Gene Ontology (GO) term enrichments associated with adjusted p values. (C) Network representation of mitotic genes significantly decreased (>2-fold change) in response to NF45 depletion. Network edges represent functional interaction between these genes.

Journal: Cell reports

Article Title: NF45 and NF90 Regulate Mitotic Gene Expression by Competing with Staufen-Mediated mRNA Decay.

doi: 10.1016/j.celrep.2020.107660

Figure Lengend Snippet: Figure 3. The NF45-NF90 Complex Regulates the Expression of a Cluster of Mitotic Genes (A) Transcriptome analysis of HepG2 cells depleted for NF45/ILF2. Red dots indicate genes that are significantly depleted by more than 2-fold change compared to control. (B) Classification of mRNAs significantly decreased in HepG2 cells depleted for NF45, based on Reactome Gene Ontology (GO) term enrichments associated with adjusted p values. (C) Network representation of mitotic genes significantly decreased (>2-fold change) in response to NF45 depletion. Network edges represent functional interaction between these genes.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NF45 Bethyl Cat# A303-147A, RRID:AB_10895652 NF90 Bethyl Cat# A303-651A, RRID:AB_11204576 NUF2 Abcam Cat# ab122962, RRID:AB_10902068 CDK1 Santa Cruz Cat# sc-54, RRID:AB_627224 CCNA2 Santa Cruz Cat# sc-53229, RRID:AB_782331 Survivin Novus Biological Cat# NB500-201, RRID:AB_10001517 Tubulin Sigma-Aldrich Cat# T5168, RRID:AB_477579 Actin Sigma-Aldrich Cat# A5441, RRID:AB_476744 Myc Millipore Cat# 05-419, RRID:AB_309725 UPF1 MBL International Cat# RN108PW, RRID:AB_2616172 p-H3 Serine 10 Cell Signaling Cat# 3377, RRID:AB_1549592 HA tag Sigma-Aldrich Cat# H6533, RRID:AB_439705 Mouse-HRP Millipore Cat# 71045-3, RRID:AB_10808067 Rabbit-HRP Millipore Cat# AP304P, RRID:AB_11212320 Tubulin Alexa Fluor 488 Thermo Fisher Scientific Cat# 322588, RRID:AB_2532182 Bacterial and Virus Strains DH5a Thermo Fisher Scientific Cat# 18265017 Experimental Models: Cell Lines HeLa ATCC Cat# CCL-2, RRID:CVCL_0030 HEK293 ATCC Cat# CRL-1573, RRID:CVCL_0045 HEK293T ATCC Cat# CRL-3216, RRID:CVCL_0063 MDA-MB-231 ATCC Cat# CRL-12532, RRID:CVCL_0062 BT-20 ATCC Cat# CRL-7912, RRID:CVCL_0178 Chemicals, Peptides, and Recombinant Proteins Nocodazole Sigma-Aldrich Cat# M1404 Thymidine Sigma-Aldrich Cat# T9250 Puromycin Sigma-Aldrich Cat# P8833 DAPI VECTASHIELD Vector laboratories Cat# H-1200 Coelenterazine-400a Biotium Cat# 10125 Polybrene Sigma-Aldrich Cat# S2667 WST-1 Sigma-Aldrich Cat# 05 015 944 001 Propidium Iodide Sigma-Aldrich Cat# P4170 RNase A Sigma-Aldrich Cat# R6513 Cycloheximide Sigma-Aldrich Biotin Thermo Fisher Scientific Cat# B1595 Tetracycline Sigma-Aldrich Cat# T7660 Hygromycin Sigma-Aldrich Cat# H3274 Protein A Sepharose GE healthcare Cat# 17-0780-01 Critical Commercial Assays RNeasy minikit Quiagen Cat# 74106 Secrete-Pair Gaussia Luciferase Assay Kit Genecopeia Cat# LF032 Luciferase Assay System Promega Cat# E1500 (Continued on next page) Cell Reports 31, 107660, May 19, 2020 e1

Techniques: Expressing, Control, Functional Assay

Figure 4. The Proximity Interactome of NF45 and NF90 Reveals a Functional Interplay with SMD Components (A) Schematic representation of the BioID pipeline. Venn diagram showing common and respective preys identified for NF45 and NF90. Dot plot of proximity interactors for NF45 and NF90 is shown. All high-confidence proximity interactors are organized according to biological functions. (B) HEK293 stably expressing inducible BirA*, BirA*-GFP, NF45-BirA*, and NF90-BirA* were induced with tetracycline and labeled with biotin during 24 h. Biotinylated proteins were precipitated using streptavidin beads, and UPF1 was detected using anti-UPF1 antibodies. (C) HEK293 cells transfected with HA-tagged Stau1 were immunoprecipitated with endogenous UPF1 using anti-UPF1 antibody. Immunoprecipitates were treated or not with RNase A (10 mg/mL). (D) qPCR of HeLa cells stably expressing shNT, shNF90, and combination of shNF90/shStau1 or shNF90/shStau2. *p < 0.05; **p < 0.01; ***p < 0.001 (two-tailed t test). (E) Relative abundance of NUF2, CDK1, CCNA2, and BIRC5 mRNAs immunoprecipitated with the anti-NF90 antibody in HEK293 cells stably expressing shNT, shStau1, or shStau2. *p < 0.05; **p < 0.01 (two-tailed t test).

Journal: Cell reports

Article Title: NF45 and NF90 Regulate Mitotic Gene Expression by Competing with Staufen-Mediated mRNA Decay.

doi: 10.1016/j.celrep.2020.107660

Figure Lengend Snippet: Figure 4. The Proximity Interactome of NF45 and NF90 Reveals a Functional Interplay with SMD Components (A) Schematic representation of the BioID pipeline. Venn diagram showing common and respective preys identified for NF45 and NF90. Dot plot of proximity interactors for NF45 and NF90 is shown. All high-confidence proximity interactors are organized according to biological functions. (B) HEK293 stably expressing inducible BirA*, BirA*-GFP, NF45-BirA*, and NF90-BirA* were induced with tetracycline and labeled with biotin during 24 h. Biotinylated proteins were precipitated using streptavidin beads, and UPF1 was detected using anti-UPF1 antibodies. (C) HEK293 cells transfected with HA-tagged Stau1 were immunoprecipitated with endogenous UPF1 using anti-UPF1 antibody. Immunoprecipitates were treated or not with RNase A (10 mg/mL). (D) qPCR of HeLa cells stably expressing shNT, shNF90, and combination of shNF90/shStau1 or shNF90/shStau2. *p < 0.05; **p < 0.01; ***p < 0.001 (two-tailed t test). (E) Relative abundance of NUF2, CDK1, CCNA2, and BIRC5 mRNAs immunoprecipitated with the anti-NF90 antibody in HEK293 cells stably expressing shNT, shStau1, or shStau2. *p < 0.05; **p < 0.01 (two-tailed t test).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NF45 Bethyl Cat# A303-147A, RRID:AB_10895652 NF90 Bethyl Cat# A303-651A, RRID:AB_11204576 NUF2 Abcam Cat# ab122962, RRID:AB_10902068 CDK1 Santa Cruz Cat# sc-54, RRID:AB_627224 CCNA2 Santa Cruz Cat# sc-53229, RRID:AB_782331 Survivin Novus Biological Cat# NB500-201, RRID:AB_10001517 Tubulin Sigma-Aldrich Cat# T5168, RRID:AB_477579 Actin Sigma-Aldrich Cat# A5441, RRID:AB_476744 Myc Millipore Cat# 05-419, RRID:AB_309725 UPF1 MBL International Cat# RN108PW, RRID:AB_2616172 p-H3 Serine 10 Cell Signaling Cat# 3377, RRID:AB_1549592 HA tag Sigma-Aldrich Cat# H6533, RRID:AB_439705 Mouse-HRP Millipore Cat# 71045-3, RRID:AB_10808067 Rabbit-HRP Millipore Cat# AP304P, RRID:AB_11212320 Tubulin Alexa Fluor 488 Thermo Fisher Scientific Cat# 322588, RRID:AB_2532182 Bacterial and Virus Strains DH5a Thermo Fisher Scientific Cat# 18265017 Experimental Models: Cell Lines HeLa ATCC Cat# CCL-2, RRID:CVCL_0030 HEK293 ATCC Cat# CRL-1573, RRID:CVCL_0045 HEK293T ATCC Cat# CRL-3216, RRID:CVCL_0063 MDA-MB-231 ATCC Cat# CRL-12532, RRID:CVCL_0062 BT-20 ATCC Cat# CRL-7912, RRID:CVCL_0178 Chemicals, Peptides, and Recombinant Proteins Nocodazole Sigma-Aldrich Cat# M1404 Thymidine Sigma-Aldrich Cat# T9250 Puromycin Sigma-Aldrich Cat# P8833 DAPI VECTASHIELD Vector laboratories Cat# H-1200 Coelenterazine-400a Biotium Cat# 10125 Polybrene Sigma-Aldrich Cat# S2667 WST-1 Sigma-Aldrich Cat# 05 015 944 001 Propidium Iodide Sigma-Aldrich Cat# P4170 RNase A Sigma-Aldrich Cat# R6513 Cycloheximide Sigma-Aldrich Biotin Thermo Fisher Scientific Cat# B1595 Tetracycline Sigma-Aldrich Cat# T7660 Hygromycin Sigma-Aldrich Cat# H3274 Protein A Sepharose GE healthcare Cat# 17-0780-01 Critical Commercial Assays RNeasy minikit Quiagen Cat# 74106 Secrete-Pair Gaussia Luciferase Assay Kit Genecopeia Cat# LF032 Luciferase Assay System Promega Cat# E1500 (Continued on next page) Cell Reports 31, 107660, May 19, 2020 e1

Techniques: Functional Assay, Stable Transfection, Expressing, Labeling, Transfection, Immunoprecipitation, Two Tailed Test