bca protein quantitation kit  (Thermo Fisher)

Journal: bioRxiv

Article Title: Targeting tRNA-Arg-TCT-4-1 suppresses cancer cell growth and tumorigenesis

doi: 10.64898/2026.02.19.706891

Figure Lengend Snippet: A) Schematic of shArg targeting sequence on tRNA-Arg-TCT-4-1. B) Northern blot showing relative tRNA-Arg-TCT-4-1 levels. Arg-TCT-4-1 lane is an overexpression construct used as a control. C) Changes in tRNA abundance upon shArg treatment. Orange circles represent tRNA-Arg-TCT isodecoders 1,2,3 and 5. D) mCherry/acGFP1 ratios measured by flow cytometry between mCherry-Hmga2-WT (5 of 12 AGA codons) and mCherry-Hmga2-MUT (0 of 12 AGA codons) reporters. Data are mean ± SD. n = 2. p values from two-way ANOVA with Šídák correction. ∗ p < 0.05 and ∗∗ p < 0.01. E) Images of 3D soft agar colony formation assays in LNZ308 cells treated with shGFP or shArg. F ) Quantification of colonies from (E). n=5, Error bars: Mean ± S.D. P values from Two-way ANOVA with posthoc Bonferroni correction. ns: not significant; ****p < 0.0001. G) In vivo tumor formation of LNZ308 cells (n = 5 or 6; error bars denote mean ± SEM). ∗∗∗∗ p < 0.0001. Two-way analysis of variance (ANOVA) and Bonferroni correction. H) Tumor weight from (G). Error bars denote mean ± SD. p value from paired Student’s t test. I) Changes in protein abundance between shArg (heavy) expressing cells and shGFP (light) control cells measured by SILAC-based proteomics; n = 3, moderated t-test. J) Codon enrichment (gene input/global genome average) of genes in cluster 2 (from ). P-value from Fisher’s Exact Test with Benjamini–Hochberg FDR correction. K) Gene Ontology enrichment analysis using downregulated proteins in cluster 2 (from ) upon tRNA-Arg-TCT-4-1 inhibition. p<0.05, FC≤1.2.

Article Snippet: LNZ308 human glioblastoma cells or LPS853 liposarcoma cells were grown in media supplemented with isotopic-labeled 13 C6 15 N2 l-lysine and 13 C6 15 N4 l-arginine (heavy) or normal amino acids (light) for 15 to 21 days until a labeling efficiency >95% was achieved following the instructions of the SILAC Protein Quantitation Kit (Trypsin) – DMEM (A33972, Thermo Scientific).

Techniques: Sequencing, Northern Blot, Over Expression, Construct, Control, Flow Cytometry, In Vivo, Quantitative Proteomics, Expressing, Multiplex sample analysis, Inhibition

Journal: bioRxiv

Article Title: Targeting tRNA-Arg-TCT-4-1 suppresses cancer cell growth and tumorigenesis

doi: 10.64898/2026.02.19.706891

Figure Lengend Snippet: A) Northern blot measuring aminoacylation levels of tRNA-Arg-TCT-4-1 in shArg vs shGFP treated LPS853 cells. B) Quantification of LPS853 tumor weight comparing shArg and shGFP control. Error bars denote mean ± SD. p value from paired Student’s t test. **p < 0.01. C) Changes in protein abundance between shArg (heavy) expressing cells and shGFP (light) control cells measured by SILAC-based proteomics; n = 3, moderated t-test. D) Codon enrichment analyses of downregulated proteins (n=576). Codon enrichment shows the fold change of each codon compared to the genome-wide average. Hierarchical agglomerative clustering, with Euclidean distance and complete linkage. E) Codon enrichment (gene input/global genome average) of genes in cluster 1 and cluster 2 ( F ). P-value from Fisher’s Exact Test with Benjamini–Hochberg FDR correction. G) Gene Ontology enrichment analysis using downregulated proteins (n=576) upon tRNA-Arg-TCT-4-1 inhibition. p<0.05, FC≤1.2.

Article Snippet: LNZ308 human glioblastoma cells or LPS853 liposarcoma cells were grown in media supplemented with isotopic-labeled 13 C6 15 N2 l-lysine and 13 C6 15 N4 l-arginine (heavy) or normal amino acids (light) for 15 to 21 days until a labeling efficiency >95% was achieved following the instructions of the SILAC Protein Quantitation Kit (Trypsin) – DMEM (A33972, Thermo Scientific).

Techniques: Northern Blot, Control, Quantitative Proteomics, Expressing, Multiplex sample analysis, Genome Wide, Inhibition

Journal: bioRxiv

Article Title: Targeting tRNA-Arg-TCT-4-1 suppresses cancer cell growth and tumorigenesis

doi: 10.64898/2026.02.19.706891

Figure Lengend Snippet: A) Northern blot measuring relative levels of tRNA-Arg-TCT-4-1 in ASO-1 treated LPS853 cells compared to untreated (UT), scramble (SCR), and mismatched (MM) controls. 20 nM, 48 hours post-transfection. B) Genomic track of the human tRNA-Arg-TCT-4-1 locus. Captured from UCSC Genome Browser. The expression level of AIM2 in ASO-1 LPS853-treated cells. (C) and shArg treatment in LPS853 and LNZ308 cells (D) . n = 3 - 4. Error bars: Mean ± S.D. P values from One-way ANOVA with posthoc Bonferroni correction. ns: not significant. E) Measurement of cell number upon treatment with ASO-1 in LP6 and 93T449 liposarcoma cell lines. 20nM, 48h after transfection. n = 3. Error bars: Mean ± S.D. P values from One-way ANOVA with posthoc Bonferroni correction. ns: not significant; ***p < 0.001; ****p < 0.0001. F) Changes in protein abundance between ASO-1 (heavy) treated cells and ASO-Ctrl (light) treated cells measured by SILAC-based proteomics; n = 3, moderated t-test. G) Gene Ontology analysis using Up or Down gene lists from ( F ).

Article Snippet: LNZ308 human glioblastoma cells or LPS853 liposarcoma cells were grown in media supplemented with isotopic-labeled 13 C6 15 N2 l-lysine and 13 C6 15 N4 l-arginine (heavy) or normal amino acids (light) for 15 to 21 days until a labeling efficiency >95% was achieved following the instructions of the SILAC Protein Quantitation Kit (Trypsin) – DMEM (A33972, Thermo Scientific).

Techniques: Northern Blot, Transfection, Expressing, Quantitative Proteomics, Multiplex sample analysis