eif4e2 antibody  (GeneTex)

Journal: iScience

Article Title: Pseudouridine synthase 1 regulates erythropoiesis via transfer RNAs pseudouridylation and cytoplasmic translation

doi: 10.1016/j.isci.2024.109265

Figure Lengend Snippet:

Article Snippet: EIF4E2 Polyclonal Antibody , ABclonal , Cat# A4305; RRID: AB_2765612.

Techniques: Recombinant, Virus, SYBR Green Assay, Cell Culture, cDNA Synthesis, Bicinchoninic Acid Protein Assay, Isolation, Iron Assay, Activity Assay, Dehydrogenase Assay, Cytochrome c Oxidase Assay, Viability Assay, RNA Sequencing, Transgenic Assay, Primer Extension Assay, Software

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 4 α1,3 fucosylated-MEST mediate interaction of MEST and eIF4E2. a Schematic of analysis strategy for identification MEST interacting protein. b GO analysis of the proteins interacted with MEST. c Venn diagrams showing the number of the proteins interact with MEST form NP, MP and GO analysis-based function. d Pull down proteins separated with SDS-PAGE and staining with CBB. e N-glycosylation of MEST at Asn163 enhances the interaction of MEST and eIF4E2. HTR-8/SVneo cells transfected with His-tagged MEST cDNA and His-tagged MEST N163Q mutant plasmid (MEST mut), respectively, then co-immunoprecipitated (co-IP) with anti His antibody and blotted with anti-eIF4E2. f FUT4 regulated interaction between MEST and eIF4E2 by regulating LeY on MEST. HTR-8/SVneo cells transfected with FUT4 cDNA and FUT4 siRNA, respectively, then interaction was detected by co-IP assay. g Confocal immunofluorescence showing the location of MEST and eIF4E2 in cells, lower panel is magnification of up panel of white frame.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: SDS Page, Staining, Glycoproteomics, Transfection, Mutagenesis, Plasmid Preparation, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 5 Genome-wide α1,3 fucosylated MEST preferentially stimulates the translation and expression of implantation relate genes. a Heatmap showing the alternation of mRNAs immunoprecipitated by anti-eIF4E2 antibody in HTR-8/SVneo cells using RIP-seq assay. HTR-8/ SVneo cells transfected with FUT4 siRNA or FUT4 cDNA. b Binding peak density analysis of binding site of mRNA and eIF4E2. c Venn diagrams displays the number of genes in FUT4 cDNA group (blue), FUT siRNA group (yellow), or both (overlapping) group. The changed of 117 overlapping mRNAs is shown in column graphs. d GO analysis of the mRNAs with high affinity to eIF4E2. e Integrative Genomics Viewer (IGV) tracks showing the binding of eIF4E2 at the 5’-UTR regions of the representative genes. f The eIF4E2-MEST/4E-T/GLGYF2 interaction is decreased in HTR-8/SVneo cells transfected with MEST mut, compared with MEST cDNA transfection. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins. g The eIF4E2-MEST/4E-T/GLGYF2 interaction in HTR-8/SVneo cells transfected with FUT4 cDNA or FUT4 siRNA, compared with scramble group. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: Genome Wide, Expressing, Immunoprecipitation, Transfection, Binding Assay, Co-Immunoprecipitation Assay, Western Blot

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 8 A summary scheme of LeY on MEST promotes embryonic trophoblast cells migration and invasion by activating translation initiation. The α1,3-fucosylation of glycoproteins in the villi of normal pregnancy and miscarriage was screened by Lectin Chip. Anti-LeY affinity chromatography was used to screen Ley scaffold glycoproteins by anti-LeY antibody. The target MEST was selected, and the specific interaction protein (eIF4E2) was discovered by anti-MEST affinity chromatography. By RIP-seq and polysome profiling, implantation-related mRNAs bound with eIF4E2 were confirmed. Reduced LeY on MEST inhibited binding of eIF4E2 with MEST, migration and invasion of trophoblast cells by inactivating translation initiation.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: Migration, Chromatography, Binding Assay

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 4 α1,3 fucosylated-MEST mediate interaction of MEST and eIF4E2. a Schematic of analysis strategy for identification MEST interacting protein. b GO analysis of the proteins interacted with MEST. c Venn diagrams showing the number of the proteins interact with MEST form NP, MP and GO analysis-based function. d Pull down proteins separated with SDS-PAGE and staining with CBB. e N-glycosylation of MEST at Asn163 enhances the interaction of MEST and eIF4E2. HTR-8/SVneo cells transfected with His-tagged MEST cDNA and His-tagged MEST N163Q mutant plasmid (MEST mut), respectively, then co-immunoprecipitated (co-IP) with anti His antibody and blotted with anti-eIF4E2. f FUT4 regulated interaction between MEST and eIF4E2 by regulating LeY on MEST. HTR-8/SVneo cells transfected with FUT4 cDNA and FUT4 siRNA, respectively, then interaction was detected by co-IP assay. g Confocal immunofluorescence showing the location of MEST and eIF4E2 in cells, lower panel is magnification of up panel of white frame.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: SDS Page, Staining, Glycoproteomics, Transfection, Mutagenesis, Plasmid Preparation, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 5 Genome-wide α1,3 fucosylated MEST preferentially stimulates the translation and expression of implantation relate genes. a Heatmap showing the alternation of mRNAs immunoprecipitated by anti-eIF4E2 antibody in HTR-8/SVneo cells using RIP-seq assay. HTR-8/ SVneo cells transfected with FUT4 siRNA or FUT4 cDNA. b Binding peak density analysis of binding site of mRNA and eIF4E2. c Venn diagrams displays the number of genes in FUT4 cDNA group (blue), FUT siRNA group (yellow), or both (overlapping) group. The changed of 117 overlapping mRNAs is shown in column graphs. d GO analysis of the mRNAs with high affinity to eIF4E2. e Integrative Genomics Viewer (IGV) tracks showing the binding of eIF4E2 at the 5’-UTR regions of the representative genes. f The eIF4E2-MEST/4E-T/GLGYF2 interaction is decreased in HTR-8/SVneo cells transfected with MEST mut, compared with MEST cDNA transfection. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins. g The eIF4E2-MEST/4E-T/GLGYF2 interaction in HTR-8/SVneo cells transfected with FUT4 cDNA or FUT4 siRNA, compared with scramble group. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: Genome Wide, Expressing, Immunoprecipitation, Transfection, Binding Assay, Co-Immunoprecipitation Assay, Western Blot

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 8 A summary scheme of LeY on MEST promotes embryonic trophoblast cells migration and invasion by activating translation initiation. The α1,3-fucosylation of glycoproteins in the villi of normal pregnancy and miscarriage was screened by Lectin Chip. Anti-LeY affinity chromatography was used to screen Ley scaffold glycoproteins by anti-LeY antibody. The target MEST was selected, and the specific interaction protein (eIF4E2) was discovered by anti-MEST affinity chromatography. By RIP-seq and polysome profiling, implantation-related mRNAs bound with eIF4E2 were confirmed. Reduced LeY on MEST inhibited binding of eIF4E2 with MEST, migration and invasion of trophoblast cells by inactivating translation initiation.

Article Snippet: After blocking in 5% non-fat dry milk for 2 h, the membranes were incubated with the primary antibody at 4 °C overnight: FUT4 (19497-1-AP), PCNA (10205-2-AP), Cyclin D1 (26939-1- AP), Cyclin E1 (11554-1-AP), MMP-2 (10373-2-AP), MMP-9 (10375-2-AP), TIMP1 (16644-1-AP), TIMP2 (17353-1-AP), MEST (11118-1-AP), eIF4E2 (12227-1-AP), GIGYF2 (24790-1-AP) and GAPDH (10494-1-AP) (Proteintech, Wuhan, China); 4E-T (sc-134233, Santa Cruz, USA); LeY (ab-3359, Abcam, USA); biotinylated LTL (B-1325-2, Vector Laboratories, California, USA).

Techniques: Migration, Chromatography, Binding Assay

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 4 α1,3 fucosylated-MEST mediate interaction of MEST and eIF4E2. a Schematic of analysis strategy for identification MEST interacting protein. b GO analysis of the proteins interacted with MEST. c Venn diagrams showing the number of the proteins interact with MEST form NP, MP and GO analysis-based function. d Pull down proteins separated with SDS-PAGE and staining with CBB. e N-glycosylation of MEST at Asn163 enhances the interaction of MEST and eIF4E2. HTR-8/SVneo cells transfected with His-tagged MEST cDNA and His-tagged MEST N163Q mutant plasmid (MEST mut), respectively, then co-immunoprecipitated (co-IP) with anti His antibody and blotted with anti-eIF4E2. f FUT4 regulated interaction between MEST and eIF4E2 by regulating LeY on MEST. HTR-8/SVneo cells transfected with FUT4 cDNA and FUT4 siRNA, respectively, then interaction was detected by co-IP assay. g Confocal immunofluorescence showing the location of MEST and eIF4E2 in cells, lower panel is magnification of up panel of white frame.

Article Snippet: Antibody eIF4E2 (12227-1-AP, Proteintech), and normal rabbit IgG (PP64B, Millipore) were used for RIP assay.

Techniques: SDS Page, Staining, Glycoproteomics, Transfection, Mutagenesis, Plasmid Preparation, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 5 Genome-wide α1,3 fucosylated MEST preferentially stimulates the translation and expression of implantation relate genes. a Heatmap showing the alternation of mRNAs immunoprecipitated by anti-eIF4E2 antibody in HTR-8/SVneo cells using RIP-seq assay. HTR-8/ SVneo cells transfected with FUT4 siRNA or FUT4 cDNA. b Binding peak density analysis of binding site of mRNA and eIF4E2. c Venn diagrams displays the number of genes in FUT4 cDNA group (blue), FUT siRNA group (yellow), or both (overlapping) group. The changed of 117 overlapping mRNAs is shown in column graphs. d GO analysis of the mRNAs with high affinity to eIF4E2. e Integrative Genomics Viewer (IGV) tracks showing the binding of eIF4E2 at the 5’-UTR regions of the representative genes. f The eIF4E2-MEST/4E-T/GLGYF2 interaction is decreased in HTR-8/SVneo cells transfected with MEST mut, compared with MEST cDNA transfection. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins. g The eIF4E2-MEST/4E-T/GLGYF2 interaction in HTR-8/SVneo cells transfected with FUT4 cDNA or FUT4 siRNA, compared with scramble group. eIF4E2 co-IP was performed, and the immunoprecipitated fractions were analyzed by immunoblotting for the indicated proteins.

Article Snippet: Antibody eIF4E2 (12227-1-AP, Proteintech), and normal rabbit IgG (PP64B, Millipore) were used for RIP assay.

Techniques: Genome Wide, Expressing, Immunoprecipitation, Transfection, Binding Assay, Co-Immunoprecipitation Assay, Western Blot

Journal: Cell death & disease

Article Title: α1,3-fucosylation of MEST promotes invasion potential of cytotrophoblast cells by activating translation initiation.

doi: 10.1038/s41419-023-06166-4

Figure Lengend Snippet: Fig. 8 A summary scheme of LeY on MEST promotes embryonic trophoblast cells migration and invasion by activating translation initiation. The α1,3-fucosylation of glycoproteins in the villi of normal pregnancy and miscarriage was screened by Lectin Chip. Anti-LeY affinity chromatography was used to screen Ley scaffold glycoproteins by anti-LeY antibody. The target MEST was selected, and the specific interaction protein (eIF4E2) was discovered by anti-MEST affinity chromatography. By RIP-seq and polysome profiling, implantation-related mRNAs bound with eIF4E2 were confirmed. Reduced LeY on MEST inhibited binding of eIF4E2 with MEST, migration and invasion of trophoblast cells by inactivating translation initiation.

Article Snippet: Antibody eIF4E2 (12227-1-AP, Proteintech), and normal rabbit IgG (PP64B, Millipore) were used for RIP assay.

Techniques: Migration, Chromatography, Binding Assay