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A ) MM lines exposed to IL-6 (100 U/ml) for 3 hrs followed by immunoblot assay for phospho-MNK and total MNK expression. Fold increase is determined by densitometric ratio of MNK-P/MNK-total and represents the mean of 3 independent experiments. * denotes IL-6-induced increase is statistically significant (p<0.05). B ) ANBL-6 cells exposed to IL-6 for 0, 60, 120 or 180 mins, followed by immunoblot assay. C ) Primary cells from 4 patients exposed to IL-6 or media (C) followed by immunoblot assay. D ) ANBL-6 cells treated with IL-6 for increasing durations, followed by immunoprecipitation with anti-MNK1, MNK2 or control IgG. Immunoprecipitates tested for phosphorylation of <t>eIF-4E</t> in vitro.

Eif 4e Full Length, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 19 article reviews

eif 4e full length - by Bioz Stars, 2026-06

93/100 stars

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1) Product Images from "MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses"

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

Journal: PLoS ONE

doi: 10.1371/journal.pone.0094011

Figure Legend Snippet: A ) MM lines exposed to IL-6 (100 U/ml) for 3 hrs followed by immunoblot assay for phospho-MNK and total MNK expression. Fold increase is determined by densitometric ratio of MNK-P/MNK-total and represents the mean of 3 independent experiments. * denotes IL-6-induced increase is statistically significant (p<0.05). B ) ANBL-6 cells exposed to IL-6 for 0, 60, 120 or 180 mins, followed by immunoblot assay. C ) Primary cells from 4 patients exposed to IL-6 or media (C) followed by immunoblot assay. D ) ANBL-6 cells treated with IL-6 for increasing durations, followed by immunoprecipitation with anti-MNK1, MNK2 or control IgG. Immunoprecipitates tested for phosphorylation of eIF-4E in vitro.

Techniques Used: Western Blot, Expressing, Immunoprecipitation, In Vitro

A ) ANBL-6 or 8226 cells transfected with empty vector (EV), serine-to-alanine phosphodefective eIF-4E (SA), serine-to-aspartic acid phosphomimetic eIF-4E (ASP) or wild type (WT) eIF-4E, followed by immunoblot assay. Transfected (transfex) vs endogenous (endog) eIF-4E denoted. B ) Above described ANBL-6 and 8226 transfected cell lines treated +/− IL-6 (100 U/ml) for varied durations and viable cell recovery enumerated. Results are relative cell growth compared to number of cells present at time 0 (mean+/−SD (n = 3)). * denotes statistically significant (p<0.05) alteration in cell recovery versus empty vector-transfected cells.

Figure Legend Snippet: A ) ANBL-6 or 8226 cells transfected with empty vector (EV), serine-to-alanine phosphodefective eIF-4E (SA), serine-to-aspartic acid phosphomimetic eIF-4E (ASP) or wild type (WT) eIF-4E, followed by immunoblot assay. Transfected (transfex) vs endogenous (endog) eIF-4E denoted. B ) Above described ANBL-6 and 8226 transfected cell lines treated +/− IL-6 (100 U/ml) for varied durations and viable cell recovery enumerated. Results are relative cell growth compared to number of cells present at time 0 (mean+/−SD (n = 3)). * denotes statistically significant (p<0.05) alteration in cell recovery versus empty vector-transfected cells.

Techniques Used: Transfection, Plasmid Preparation, Western Blot

A ) 8226 cells injected SQ and mice randomized to control (C) group, low dose CGP57380 (LD, 20 mg/kg) or high dose (HD, 50 mg/kg). Treatment is daily IP injections. At day +4 and +7 of treatment, tumors were measured. Data is tumor volume, mean+/−SD (n = 8). * = significantly (p<0.05) smaller tumor vs control. B and C ) Individual control (C), high dose (HD)-treated or low dose (LD)-treated tumors harvested after 3 days of treatment for Western blot, assaying phospho-eIF-4E, total eIF-4E, GAPDH and PARP that has been cleaved by caspase (AB #954 from Cell Signaling). D ) Mice challenged SQ with 8226 cells transfected with EV (right flank) and 8226 cells transfected with dominant negative eIF-4E (left flank). A separate cohort of mice were challenged with non-transfected parental 8226 cells. Tumor growth is mean+/−SD tumor volume, n = 8.

Figure Legend Snippet: A ) 8226 cells injected SQ and mice randomized to control (C) group, low dose CGP57380 (LD, 20 mg/kg) or high dose (HD, 50 mg/kg). Treatment is daily IP injections. At day +4 and +7 of treatment, tumors were measured. Data is tumor volume, mean+/−SD (n = 8). * = significantly (p<0.05) smaller tumor vs control. B and C ) Individual control (C), high dose (HD)-treated or low dose (LD)-treated tumors harvested after 3 days of treatment for Western blot, assaying phospho-eIF-4E, total eIF-4E, GAPDH and PARP that has been cleaved by caspase (AB #954 from Cell Signaling). D ) Mice challenged SQ with 8226 cells transfected with EV (right flank) and 8226 cells transfected with dominant negative eIF-4E (left flank). A separate cohort of mice were challenged with non-transfected parental 8226 cells. Tumor growth is mean+/−SD tumor volume, n = 8.

Techniques Used: Injection, Western Blot, Transfection, Dominant Negative Mutation

A ) AHA incorporation is tested in MM cells stimulated with IL-6 for 30, 60, 120 or 180 mins. In B ), AHA incorporation is tested in IL-6-stimulated EV-, WT or mutant SA eIF-4E-transfected MM cells. In C ), EV or SA mutant-expressing cells are pre-treated +/− pp242, followed by IL-6. Bar graphs under panels are means+/−SD of 4 separate experiments. * denotes statistically significant increase in AHA incorporation induced by IL-6 in A and B and decrease versus IL-6-stimulated control in C). D ) Lambda light chain ELISA assay in MM cell lysates, comparing EV- vs mutant eIF-4E-transfected cells +/− IL-6 and +/− pp242. Data are means+/−SD, n = 4. * denotes statistically significant difference from corresponding controls.

Figure Legend Snippet: A ) AHA incorporation is tested in MM cells stimulated with IL-6 for 30, 60, 120 or 180 mins. In B ), AHA incorporation is tested in IL-6-stimulated EV-, WT or mutant SA eIF-4E-transfected MM cells. In C ), EV or SA mutant-expressing cells are pre-treated +/− pp242, followed by IL-6. Bar graphs under panels are means+/−SD of 4 separate experiments. * denotes statistically significant increase in AHA incorporation induced by IL-6 in A and B and decrease versus IL-6-stimulated control in C). D ) Lambda light chain ELISA assay in MM cell lysates, comparing EV- vs mutant eIF-4E-transfected cells +/− IL-6 and +/− pp242. Data are means+/−SD, n = 4. * denotes statistically significant difference from corresponding controls.

Techniques Used: Mutagenesis, Transfection, Expressing, Enzyme-linked Immunosorbent Assay

A ) Different categories of 166 genes demonstrating significantly inhibited translation in mutant-expressing cells. B ) Immunoblot analysis of protein expression in EV-, WT and mutant (MU)-expressing MM cells. C ) Immunoblot analysis of protein expression in EV vs mutant (MU) eIF-4E-expressing MM cells +/− IL-6 for 48 hrs.

Figure Legend Snippet: A ) Different categories of 166 genes demonstrating significantly inhibited translation in mutant-expressing cells. B ) Immunoblot analysis of protein expression in EV-, WT and mutant (MU)-expressing MM cells. C ) Immunoblot analysis of protein expression in EV vs mutant (MU) eIF-4E-expressing MM cells +/− IL-6 for 48 hrs.

Techniques Used: Mutagenesis, Expressing, Western Blot

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Journal: PLoS ONE

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

doi: 10.1371/journal.pone.0094011

Figure Lengend Snippet: A ) MM lines exposed to IL-6 (100 U/ml) for 3 hrs followed by immunoblot assay for phospho-MNK and total MNK expression. Fold increase is determined by densitometric ratio of MNK-P/MNK-total and represents the mean of 3 independent experiments. * denotes IL-6-induced increase is statistically significant (p<0.05). B ) ANBL-6 cells exposed to IL-6 for 0, 60, 120 or 180 mins, followed by immunoblot assay. C ) Primary cells from 4 patients exposed to IL-6 or media (C) followed by immunoblot assay. D ) ANBL-6 cells treated with IL-6 for increasing durations, followed by immunoprecipitation with anti-MNK1, MNK2 or control IgG. Immunoprecipitates tested for phosphorylation of eIF-4E in vitro.

Article Snippet: The HA-tagged eIF-4E full length coding sequence was isolated from pHA-eIF4E (obtained from Addgene, plasmid 17343) by PCR.

Techniques: Western Blot, Expressing, Immunoprecipitation, In Vitro

Journal: PLoS ONE

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

doi: 10.1371/journal.pone.0094011

Figure Lengend Snippet: A ) ANBL-6 or 8226 cells transfected with empty vector (EV), serine-to-alanine phosphodefective eIF-4E (SA), serine-to-aspartic acid phosphomimetic eIF-4E (ASP) or wild type (WT) eIF-4E, followed by immunoblot assay. Transfected (transfex) vs endogenous (endog) eIF-4E denoted. B ) Above described ANBL-6 and 8226 transfected cell lines treated +/− IL-6 (100 U/ml) for varied durations and viable cell recovery enumerated. Results are relative cell growth compared to number of cells present at time 0 (mean+/−SD (n = 3)). * denotes statistically significant (p<0.05) alteration in cell recovery versus empty vector-transfected cells.

Article Snippet: The HA-tagged eIF-4E full length coding sequence was isolated from pHA-eIF4E (obtained from Addgene, plasmid 17343) by PCR.

Techniques: Transfection, Plasmid Preparation, Western Blot

Journal: PLoS ONE

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

doi: 10.1371/journal.pone.0094011

Figure Lengend Snippet: A ) 8226 cells injected SQ and mice randomized to control (C) group, low dose CGP57380 (LD, 20 mg/kg) or high dose (HD, 50 mg/kg). Treatment is daily IP injections. At day +4 and +7 of treatment, tumors were measured. Data is tumor volume, mean+/−SD (n = 8). * = significantly (p<0.05) smaller tumor vs control. B and C ) Individual control (C), high dose (HD)-treated or low dose (LD)-treated tumors harvested after 3 days of treatment for Western blot, assaying phospho-eIF-4E, total eIF-4E, GAPDH and PARP that has been cleaved by caspase (AB #954 from Cell Signaling). D ) Mice challenged SQ with 8226 cells transfected with EV (right flank) and 8226 cells transfected with dominant negative eIF-4E (left flank). A separate cohort of mice were challenged with non-transfected parental 8226 cells. Tumor growth is mean+/−SD tumor volume, n = 8.

Article Snippet: The HA-tagged eIF-4E full length coding sequence was isolated from pHA-eIF4E (obtained from Addgene, plasmid 17343) by PCR.

Techniques: Injection, Western Blot, Transfection, Dominant Negative Mutation

Journal: PLoS ONE

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

doi: 10.1371/journal.pone.0094011

Figure Lengend Snippet: A ) AHA incorporation is tested in MM cells stimulated with IL-6 for 30, 60, 120 or 180 mins. In B ), AHA incorporation is tested in IL-6-stimulated EV-, WT or mutant SA eIF-4E-transfected MM cells. In C ), EV or SA mutant-expressing cells are pre-treated +/− pp242, followed by IL-6. Bar graphs under panels are means+/−SD of 4 separate experiments. * denotes statistically significant increase in AHA incorporation induced by IL-6 in A and B and decrease versus IL-6-stimulated control in C). D ) Lambda light chain ELISA assay in MM cell lysates, comparing EV- vs mutant eIF-4E-transfected cells +/− IL-6 and +/− pp242. Data are means+/−SD, n = 4. * denotes statistically significant difference from corresponding controls.

Article Snippet: The HA-tagged eIF-4E full length coding sequence was isolated from pHA-eIF4E (obtained from Addgene, plasmid 17343) by PCR.

Techniques: Mutagenesis, Transfection, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: MNK1-Induced eIF-4E Phosphorylation in Myeloma Cells: A Pathway Mediating IL-6-Induced Expansion and Expression of Genes Involved in Metabolic and Proteotoxic Responses

doi: 10.1371/journal.pone.0094011

Figure Lengend Snippet: A ) Different categories of 166 genes demonstrating significantly inhibited translation in mutant-expressing cells. B ) Immunoblot analysis of protein expression in EV-, WT and mutant (MU)-expressing MM cells. C ) Immunoblot analysis of protein expression in EV vs mutant (MU) eIF-4E-expressing MM cells +/− IL-6 for 48 hrs.

Article Snippet: The HA-tagged eIF-4E full length coding sequence was isolated from pHA-eIF4E (obtained from Addgene, plasmid 17343) by PCR.

Techniques: Mutagenesis, Expressing, Western Blot

Figure 3. Distribution of eIF4E between free and complexed (4EBP1 and eIF4G) forms in mammary parenchyma of nonpregnant lactating (white bars) and nonlactating (hatched bars) cows. Data are means ± SE for 3 animals/group. For each variable, values not sharing the same letter are signiﬁcantly different, P ≤0.05.

Journal: Journal of dairy science

Article Title: Abundance and phosphorylation state of translation initiation factors in mammary glands of lactating and nonlactating dairy cows.

doi: 10.3168/jds.2006-778

Figure Lengend Snippet: Figure 3. Distribution of eIF4E between free and complexed (4EBP1 and eIF4G) forms in mammary parenchyma of nonpregnant lactating (white bars) and nonlactating (hatched bars) cows. Data are means ± SE for 3 animals/group. For each variable, values not sharing the same letter are signiﬁcantly different, P ≤0.05.

Article Snippet: Unphosphorylated and phosphorylated eIF4E were detected, after separation on 12% gels, with a polyclonal anti-eIF4E IgG raised against a recombinant protein representing the full-length eIF4E of human origin (Santa Cruz Biotechnology, Inc.), and a polyclonal antibody against a synthetic phosphopeptide corresponding to residues surrounding Ser209 of human eIF4E (Cell Signaling).

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