rabbit eif4e  (Novus Biologicals)

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