Journal: bioRxiv
Article Title: tRF-3021a, a tRNA-Ala-TGC derived 3’ fragment, promotes glioblastoma cell invasion, suppresses apoptosis, and is required for normal levels of protein synthesis
doi: 10.64898/2026.01.26.701835
Figure Lengend Snippet: (A) Time course quantification of apoptotic (Annexin V–positive) U251 cells after transfection with negative control (NC) LNA or tRF-3021a LNA at the indicated time points. (B) Immunoblot time course of cleaved PARP in U251 cells following tRF-3021a knockdown (KD) versus NC LNA; GAPDH serves as a loading control. (C) Puromycin -labeling time course in U251 cells following tRF-3021a KD versus NC LNA. Cells were incubated with puromycin, and lysates were immunoblotted with anti-puromycin to assess global protein synthesis; GAPDH serves as a loading control. (D) Time course immunoblot analysis (2–12 h) of translation-regulatory signaling following tRF-3021a KD versus NC LNA, including total and phospho-eIF2α, total and phospho-p70S6 kinase (Thr389), and phospho-4E-BP1 (Thr37/46); GAPDH serves as a loading control. (E) Quantification of apoptotic (Annexin V–positive) cells after tRF-3021a KD in the presence or absence of the pan-caspase inhibitor z-VAD-fmk. (F) Puromycin-labeling assay at 48 h in U251 cells transfected with NC LNA, tRF-3021a LNA, tRF-3021a LNA + z-VAD-fmk or treated with z-VAD-fmk alone. Statistics: multiple unpaired two-tailed t tests comparing KD to NC at each time point with Benjamini–Krieger–Yekutieli false-discovery-rate correction (Q = 1%); p = 0.007751 (12 h), p = 0.000205 (24 h), p < 0.000001 (48 h). (E) one-way ANOVA with Tukey’s multiple-comparisons test; p = 0.0316 for KD versus KD + z-VAD. Significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
Article Snippet: For chemical treatment, cells were treated with the pan-caspase inhibitor z-VAD-fmk (InvivoGen, tlrl-vad) at a final concentration of 20 μM for 48 h. To maintain inhibition, fresh z-VAD-fmk was added again at 24 h.
Techniques: Transfection, Negative Control, Western Blot, Knockdown, Control, Labeling, Incubation, Two Tailed Test
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