eif4e  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Selective targeting of type II tRNAs underlies SLFN14-mediated translational repression and its dysregulation by thrombocytopenia-linked mutations

doi: 10.64898/2026.02.03.703619

Figure Lengend Snippet: (A) 293T cells were induced to express SLFN14 for 6, 12, or 24 h, and cell viability was assessed using the MTT assay. (B) Western blot analysis of mCherry-SLFN14, HSP70, β-actin, and GAPDH expression. (C) Western blot analysis of mTORC1 pathway markers. (D) Western blot analysis of total eIF4E and phosphorylated eIF4E. (E) Schematic model illustrating how the IT-linked K219N mutation disrupts protein synthesis by shifting SLFN14 RNA cleavage activity toward type II tRNAs, leading to ribosome stalling, activation of compensatory stress pathways, and ultimately cell death.

Article Snippet: The antibodies against the following proteins were used in the immunoblotting: mCherry (Bio-techne, NBP2-25157), HSP70 (Santa Cruz, sc-137210), β-actin (Sigma, A2228), GAPDH (Cell Signaling Technology, #2118), 4E-BP1 (Cell Signaling Technology, #9452), Phospho-4E-BP1(Thr37/46) (Cell Signaling Technology, #2855), eIF4E (Cell Signaling Technology, #9742), Phospho-eIF4E (Ser209) (Cell Signaling Technology, #9741), p70 S6 Kinase (Cell Signaling Technology, #9202), Phospho-p70 S6 Kinase (Cell Signaling Technology, #9205).

Techniques: MTT Assay, Western Blot, Expressing, Mutagenesis, Activity Assay, Activation Assay

Journal: Oncogene

Article Title: Identification of Raptor and GLI1 as USP37 substrates highlight its context-specific function in medulloblastoma cells

doi: 10.1038/s41388-025-03651-2

Figure Lengend Snippet: A Western blotting to show changes in the levels of Raptor, 4EBP1, p-4EBP1 and p27 in DAOY and ONS76 MB cell lines with constitutive over expression of USP37 . B Schema to illustrate expected effect of mTORC1 kinase activity on phosphorylation of 4EBP1 and its interaction with eIF4E. Phosphorylated 4EBP1 releases eIF4E, allowing its binding to the CAP structure of mRNA and thereby enabling canonical protein translation. Conversely, mTORC1 inactivity results in the binding of unphosphorylated 4EBP1 to eIF4E, preventing its association with the CAP structure of mRNA. This is expected to prevent canonical protein translation initiation and cause a potential shift to CAP-independent protein translation. Co-immunoprecipitation assays to study C interaction of 4EBP1 with eIF4E and eIF4G using control IgG or anti-4EBP1 antibody and D eIF4E with 4EBP1, p-4EBP1, and eIF4G using control IgG or anti-eIF4E antibody, in parental and isogenic USP37- and REST- overexpressing DAOY and ONS76 cells, respectively. Statistical data are presented for three independent biological replicates as the means ± SDs. ns = non-significant, *p < 0.05, **p < 0.01, and ****p < 0.0001 by Student’s t test.

Article Snippet: Lysates were incubated with control mouse IgG, anti-USP37 (Cat# A300-927A, Thermo Fisher Scientific), anti-4EBP1 (Cat# 9644, Cell Signaling Technology, MA, USA), anti-eIF4E (Cat# 2067, Cell Signaling Technology) primary antibodies overnight at 4 °C and then incubated with PierceTM Protein A/G UltraLinkTM Resin (Cat# 53132, Thermo Fisher Scientific) for 1.5 h at 4 °C.

Techniques: Western Blot, Over Expression, Activity Assay, Phospho-proteomics, Binding Assay, Immunoprecipitation, Control