Review



adherent hek293t cells  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 99
      Buy from Supplier

    Structured Review

    ATCC adherent hek293t cells
    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. <t>HEK293T</t> 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Adherent Hek293t Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 38021 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/adherent+hek293t+cells/293T/bio_rxiv__64898__2026__05__26__727988-171-0-4
    Average 99 stars, based on 38021 article reviews
    adherent hek293t cells - by Bioz Stars, 2026-09
    99/100 stars

    Images

    1) Product Images from "Eukaryotic tRNA ligases mediate RNA break repair"

    Article Title: Eukaryotic tRNA ligases mediate RNA break repair

    Journal: bioRxiv

    doi: 10.64898/2026.05.26.727988

    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. HEK293T 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Figure Legend Snippet: (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. HEK293T 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Techniques Used: Northern Blot, Positive Control, Purification, Incubation, Control, Luciferase, In Vitro, Isolation, Activity Assay, Two Tailed Test, Standard Deviation

    (A) Left: Schematic of the direct RNA nanopore sequencing for the restrictocin cleaved and RTCB/Trl1 repaired ribosomes. Right: Schematic of sequence coverage plots of the 28S rRNA around the SRL cleavage site. After cleavage with restrictocin, the sequence coverage drops, this drop is reduced after break repair with both tRNA ligases. (B) The sequence coverage drop is reduced by repair with both tRNA ligases. The sequence coverage 5′ of the cleavage site (nt 4550) was normalized to the sequence coverage 3′ of the cleavage site (nt 4650) for all analyzed samples after nanopore sequencing. n=4 independent experiments, statistical significance of cleavage and repair was determined by One-way ANOVA followed by Tukey’s multiple comparison test. (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***). (C) Around 60% of cleaved HEK293T ribosomes were repaired by RTCB, whereas ∼70% of ribosomes were repaired by Trl1. (D) Schematic of the unspecific breakage and repair assay, using RNase 4, which cleaves between any accessible U/A, or U/G dinucleotide on the isolated ribosomes, followed by repair with RTCB or Trl1 and nanopore sequencing-based breakage and repair site-mapping. (E-H) Most RNase4-induced RNA breaks show reduced relative 5′-end peak intensity or 3′-end peak intensity for both 28S and 18S rRNA after repair with RTCB (n=3 independent experiments). Hits were selected, when present in at least two replicates and when found by both analyses (<30 nt between 5′-end and 3′-end peak intensity). (I) Structural snapshots of the human ribosome (PDB 4ug0) . Shown in dark violet is the SRL as a reference point, the breakage sites identified by either analysis are shown in dark red, the space in between the mapped break sites is shown in light green. Error bars depict standard deviation (SD).
    Figure Legend Snippet: (A) Left: Schematic of the direct RNA nanopore sequencing for the restrictocin cleaved and RTCB/Trl1 repaired ribosomes. Right: Schematic of sequence coverage plots of the 28S rRNA around the SRL cleavage site. After cleavage with restrictocin, the sequence coverage drops, this drop is reduced after break repair with both tRNA ligases. (B) The sequence coverage drop is reduced by repair with both tRNA ligases. The sequence coverage 5′ of the cleavage site (nt 4550) was normalized to the sequence coverage 3′ of the cleavage site (nt 4650) for all analyzed samples after nanopore sequencing. n=4 independent experiments, statistical significance of cleavage and repair was determined by One-way ANOVA followed by Tukey’s multiple comparison test. (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***). (C) Around 60% of cleaved HEK293T ribosomes were repaired by RTCB, whereas ∼70% of ribosomes were repaired by Trl1. (D) Schematic of the unspecific breakage and repair assay, using RNase 4, which cleaves between any accessible U/A, or U/G dinucleotide on the isolated ribosomes, followed by repair with RTCB or Trl1 and nanopore sequencing-based breakage and repair site-mapping. (E-H) Most RNase4-induced RNA breaks show reduced relative 5′-end peak intensity or 3′-end peak intensity for both 28S and 18S rRNA after repair with RTCB (n=3 independent experiments). Hits were selected, when present in at least two replicates and when found by both analyses (<30 nt between 5′-end and 3′-end peak intensity). (I) Structural snapshots of the human ribosome (PDB 4ug0) . Shown in dark violet is the SRL as a reference point, the breakage sites identified by either analysis are shown in dark red, the space in between the mapped break sites is shown in light green. Error bars depict standard deviation (SD).

    Techniques Used: Nanopore Sequencing, Sequencing, Comparison, Isolation, Standard Deviation

    (A) Schematic of the experimental setup treating the RTCB knockdown cells with increasing concentrations of menadione for 3 h, before analyzing the isolated total cellular via capillary electrophoresis. (B) Electropherograms showing the effect of the RTCB knockdown with increasing concentrations of menadione. With increasing menadione concentrations, RNA is more fragmented in the RTCB KD compared to the WT. (C) Plots for the relative fluorescent units (a.u., arbitrary units) for three concentrations of menadione (0 µM, 60 µM and 100 µM) of the electropherograms shown in (B). Top: Whole cell RNA plot. Bottom: zoom in to small RNA fragments (nt 0-1500). (D) eCDF plots of cumulative probability for each read length after nanopore sequencing of RNA from cells treated with 0 µM and 60 µM menadione. Shown are read lengths for 28S rRNA (axis break between 2999 nt and 4500 nt) and 18S. (E) eCDF plots of cumulative probability for each read length after nanopore sequencing using specific sequencing adapters for RPPH1 and RN7SK RNA from cells treated with 0 µM and 60 µM menadione. (F) Puromycin incorporation assay for HEK293T cells, treated with harringtonine as a control, and 3 different concentrations of menadione (40 µM, 60 µM, 100 µM) for 1 h. Median fluorescence intensity (MFI) of puromycin-AF488 is shown. For gating strategy refer to Figure S5C. (G) Polysome profiling (15-50% sucrose gradient fractionation) using either untreated cells, or cells treated with 60 µM menadione for 1 h. For quantification, profiles are categorized in three sections: 1) free mRNPs and 40S subunits, 2) sub- and light polysomes and 3) heavy polysomes. The peaks for 40S and 60S subunits, as well as for 80S ribosomes and polysomes are indicated. (H) Quantification of the AUC of the polysome profiles of ribosome/total ratios according to the 3 categories. N=4 independent experiments. (I) Western blot analysis specific for RTCB, and the ribosomal proteins RPLS6 (40S) and RPL10A (60S) using sucrose fractions from (H). (J) Viability assay of RTCB WT and RTCB knockdown cells treated with increasing concentrations of menadione. With increasing concentrations of menadione, cells are less viable in the RTCB KD compared to the WT. n=7 biological replicates. (K) Area under the curve (AUC) for the viability plots in (F). The AUC for the viability in the KD cells in significantly lower compared to the WT. Statistical significances for all panels in this figure were determined using unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Figure Legend Snippet: (A) Schematic of the experimental setup treating the RTCB knockdown cells with increasing concentrations of menadione for 3 h, before analyzing the isolated total cellular via capillary electrophoresis. (B) Electropherograms showing the effect of the RTCB knockdown with increasing concentrations of menadione. With increasing menadione concentrations, RNA is more fragmented in the RTCB KD compared to the WT. (C) Plots for the relative fluorescent units (a.u., arbitrary units) for three concentrations of menadione (0 µM, 60 µM and 100 µM) of the electropherograms shown in (B). Top: Whole cell RNA plot. Bottom: zoom in to small RNA fragments (nt 0-1500). (D) eCDF plots of cumulative probability for each read length after nanopore sequencing of RNA from cells treated with 0 µM and 60 µM menadione. Shown are read lengths for 28S rRNA (axis break between 2999 nt and 4500 nt) and 18S. (E) eCDF plots of cumulative probability for each read length after nanopore sequencing using specific sequencing adapters for RPPH1 and RN7SK RNA from cells treated with 0 µM and 60 µM menadione. (F) Puromycin incorporation assay for HEK293T cells, treated with harringtonine as a control, and 3 different concentrations of menadione (40 µM, 60 µM, 100 µM) for 1 h. Median fluorescence intensity (MFI) of puromycin-AF488 is shown. For gating strategy refer to Figure S5C. (G) Polysome profiling (15-50% sucrose gradient fractionation) using either untreated cells, or cells treated with 60 µM menadione for 1 h. For quantification, profiles are categorized in three sections: 1) free mRNPs and 40S subunits, 2) sub- and light polysomes and 3) heavy polysomes. The peaks for 40S and 60S subunits, as well as for 80S ribosomes and polysomes are indicated. (H) Quantification of the AUC of the polysome profiles of ribosome/total ratios according to the 3 categories. N=4 independent experiments. (I) Western blot analysis specific for RTCB, and the ribosomal proteins RPLS6 (40S) and RPL10A (60S) using sucrose fractions from (H). (J) Viability assay of RTCB WT and RTCB knockdown cells treated with increasing concentrations of menadione. With increasing concentrations of menadione, cells are less viable in the RTCB KD compared to the WT. n=7 biological replicates. (K) Area under the curve (AUC) for the viability plots in (F). The AUC for the viability in the KD cells in significantly lower compared to the WT. Statistical significances for all panels in this figure were determined using unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Techniques Used: Knockdown, Isolation, Electrophoresis, Nanopore Sequencing, Sequencing, Control, Fluorescence, Fractionation, Western Blot, Viability Assay, Standard Deviation

    Related Articles

    Cell Culture:

    Article Title: Cell-type-focused compound screen in human organoids reveals CK1 inhibition protects cone photoreceptors from death.
    Article Snippet: .. Adherent HEK293T cells (ATCC, #CRL3216) were cultured in a 5-layer CellSTACK (3,180 cm2; Corning, # CLS3319) for AAV vector production. ..

    Article Title: Triple Labeling Resolves a GPCR Intermediate State by Using Three-Color Single Molecule FRET
    Article Snippet: .. All expressions were carried out using adherent HEK293T cells (American Type Culture Collection CRL-3216, LGC Standards S.a.r.l., France) cultured in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich) at 37 °C with 5% CO 2 . .. To improve adherence, all culture vessels were treated with a 28.5 μg/μL solution of poly- l -ornithine-hydrobromide (Sigma) in DPBS at 37 °C for 15 min followed by a single wash using DPBS before cell plating.

    Bioprocessing:

    Article Title: Cell-type-focused compound screen in human organoids reveals CK1 inhibition protects cone photoreceptors from death.
    Article Snippet: .. Adherent HEK293T cells (ATCC, #CRL3216) were cultured in a 5-layer CellSTACK (3,180 cm2; Corning, # CLS3319) for AAV vector production. ..

    Plasmid Preparation:

    Article Title: Cell-type-focused compound screen in human organoids reveals CK1 inhibition protects cone photoreceptors from death.
    Article Snippet: .. Adherent HEK293T cells (ATCC, #CRL3216) were cultured in a 5-layer CellSTACK (3,180 cm2; Corning, # CLS3319) for AAV vector production. ..

    Modification:

    Article Title: Eukaryotic tRNA ligases mediate RNA break repair
    Article Snippet: RNA samples were unfolded at 80°C for 2 min, analyzed by 15% denaturing urea-PAGE and visualized via SYBRTM Gold nucleic acid gel stain (Invitrogen, Life Technologies). .. Adherent HEK293T cells (CRL-3216, American Tissue Culture Collection, ATCC) (kind gift from the Christina Paulino lab) were grown at 37°C, 5% carbon dioxide and 70% humidity in 10cm dishes (Gibco) in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and antibiotic/antimycotic solution (Capricorn Scientific). .. In the Leppek lab, human HEK393T (ATCC: CRL-3216) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco) containing 2 mM L-glutamine, supplemented with 10% fetal calf serum (Gibco), 100 U/ml penicillin and 0.1 mg/mL streptomycin (EMD Millipore, or Gibco,) (regular medium, RM) at 37°C in 5% CO 2 -buffered incubators.

    Article Title: Chronologically distributed transfection improves AAV2 and AAV2/8 capsid filling and reveals assembly schedule divergence
    Article Snippet: FreeStyle HEK293-F cells (Life Technologies Limited, Paisley, UK, catalog number R79007 ) were cultivated in FreeStyle 293 Expression Medium (Life Technologies, cat. no. 12338018) at 37°C with 5% CO 2 and shaken at 135 rpm inside a humidified incubator (PHC Europe B.V., Loughborough, UK, model MCO-170AICD-PE). .. Adherent HEK293T cells (cat. no. CRL-3216) from American type culture collection (ATCC) were cultivated using Dulbecco’s modified Eagle’s medium (DMEM) with 10% v/v heat-inactivated fetal bovine serum (FBS), both purchased from Gibco, UK. .. Cells were kept at 37°C, 5% CO 2 also in a PHC Europe, model MCO-170AICD-PE, humidified incubator.

    Article Title: Triple Labeling Resolves a GPCR Intermediate State by Using Three-Color Single Molecule FRET
    Article Snippet: .. All expressions were carried out using adherent HEK293T cells (American Type Culture Collection CRL-3216, LGC Standards S.a.r.l., France) cultured in Dulbecco’s modified Eagle’s medium (DMEM; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich) at 37 °C with 5% CO 2 . .. To improve adherence, all culture vessels were treated with a 28.5 μg/μL solution of poly- l -ornithine-hydrobromide (Sigma) in DPBS at 37 °C for 15 min followed by a single wash using DPBS before cell plating.

    Patch Clamp:

    Article Title: Structural Mechanism of Prestin-Membrane Mechanotransduction
    Article Snippet: These cells were grown at 37°C and 7.8% CO 2 in FreeStyleTM 293 expression medium (Gibco Fisher Scientific) supplemented with 10 μg/ml penicillin/streptomycin (Company) and 2% heat-inactivated fetal bovine serum (FBS) (Company). .. Adherent HEK293T cells were obtained from ATCC (ATCC CRL-3216) and used for patch-clamp electrophysiology experiments. .. They were cultured at 37°C and 5% CO 2 in in Dulbecco’s Modified Eagle’s Media (DMEM, Gibco Fisher Scientific) supplemented with 10% heat-inactivated FBS.

    Expressing:

    Article Title: Multivalent display of membrane proteins using HIV-1 Gag VLPs
    Article Snippet: .. Adaptation to serum-free media of HEK293T The adherent HEK293T cells (ATCC, CRL-3216) grown in DMEM (Gibco, 10566-016) + 10% FBS were subjected to gradually reducing concentrations of FBS, reducing 5% to 2.5% to 1.25% in DMEM, which were finally transitioned to FreeStyle 293 expression media (Thermo Fisher Scientific, catalog #12338018). ..



    Similar Products

    adherent hek293t cells  (ATCC)
    99
    ATCC adherent hek293t cells
    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. <t>HEK293T</t> 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Adherent Hek293t Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/adherent+hek293t+cells/293T/bio_rxiv__64898__2026__05__26__727988-171-0-4
    Average 99 stars, based on 1 article reviews
    adherent hek293t cells - by Bioz Stars, 2026-09
    99/100 stars
    		  Buy from Supplier
    adherent hek293t cells  (Mirus Bio)
    97
    Mirus Bio adherent hek293t cells
    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. <t>HEK293T</t> 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Adherent Hek293t Cells, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/adherent+hek293t+cells/TransIT+-293/bio_rxiv__64898__2026__04__16__718981-190-8-16
    Average 97 stars, based on 1 article reviews
    adherent hek293t cells - by Bioz Stars, 2026-09
    97/100 stars
    		  Buy from Supplier
    c adherent hek293t cells  (Santa Cruz Biotechnology)
    94
    Santa Cruz Biotechnology c adherent hek293t cells
    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. <t>HEK293T</t> 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    C Adherent Hek293t Cells, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/adherent+hek293t+cells/HEK293T+Whole+Cell+Lysate/pmc10897633__mmc2-241-14-38
    Average 94 stars, based on 1 article reviews
    c adherent hek293t cells - by Bioz Stars, 2026-09
    94/100 stars
    		  Buy from Supplier
    adherent hek293t cell line  (ATCC)
    99
    ATCC adherent hek293t cell line
    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. <t>HEK293T</t> 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).
    Adherent Hek293t Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/adherent+hek293t+cells/293T/pmc10928139__mmc2-255-0-4
    Average 99 stars, based on 1 article reviews
    adherent hek293t cell line - by Bioz Stars, 2026-09
    99/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. HEK293T 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Journal: bioRxiv

    Article Title: Eukaryotic tRNA ligases mediate RNA break repair

    doi: 10.64898/2026.05.26.727988

    Figure Lengend Snippet: (A) Schematic of the density-ultracentrifugation assay to investigate whether the α-fragment remains attached to the ribosome after cleavage with restrictocin. (B) The α-fragment remains attached to the ribosome, by Northern blot it is only detected in the positive control and in the ribosome-containing pellet fraction. n=3 independent experiments. (C) Urea-PAGE analysis of 80S ribosome cleavage and repair. HEK293T 80S ribosomes are cleaved with purified restrictocin, after cleavage the α-fragment band appears, which is reduced in intensity after incubation with RTCB or Trl1 for 2 h. (D) Quantification of (C). Repair with both tRNA ligases leads to a significant decrease in α-fragment-band intensity, normalized to 5S rRNA as a loading control. n=4 independent experiments. (E) Schematic for the hybrid luciferase in vitro translation assay, using ribosome-depleted RRL and isolated 80S ribosomes from HEK293T cells to assay for translational activity after SRL cleavage and repair. (F) Luciferase activity measurement of hybrid luciferase in vitro translation assay. Repair of inactivated ribosomes with RTCB significantly increases translational activity in the hybrid luciferase in vitro translation assay. n=3 independent experiments. Statistical significances for panels in this figure were determined using two tailed, unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Article Snippet: Adherent HEK293T cells (CRL-3216, American Tissue Culture Collection, ATCC) (kind gift from the Christina Paulino lab) were grown at 37°C, 5% carbon dioxide and 70% humidity in 10cm dishes (Gibco) in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and antibiotic/antimycotic solution (Capricorn Scientific).

    Techniques: Northern Blot, Positive Control, Purification, Incubation, Control, Luciferase, In Vitro, Isolation, Activity Assay, Two Tailed Test, Standard Deviation

    (A) Left: Schematic of the direct RNA nanopore sequencing for the restrictocin cleaved and RTCB/Trl1 repaired ribosomes. Right: Schematic of sequence coverage plots of the 28S rRNA around the SRL cleavage site. After cleavage with restrictocin, the sequence coverage drops, this drop is reduced after break repair with both tRNA ligases. (B) The sequence coverage drop is reduced by repair with both tRNA ligases. The sequence coverage 5′ of the cleavage site (nt 4550) was normalized to the sequence coverage 3′ of the cleavage site (nt 4650) for all analyzed samples after nanopore sequencing. n=4 independent experiments, statistical significance of cleavage and repair was determined by One-way ANOVA followed by Tukey’s multiple comparison test. (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***). (C) Around 60% of cleaved HEK293T ribosomes were repaired by RTCB, whereas ∼70% of ribosomes were repaired by Trl1. (D) Schematic of the unspecific breakage and repair assay, using RNase 4, which cleaves between any accessible U/A, or U/G dinucleotide on the isolated ribosomes, followed by repair with RTCB or Trl1 and nanopore sequencing-based breakage and repair site-mapping. (E-H) Most RNase4-induced RNA breaks show reduced relative 5′-end peak intensity or 3′-end peak intensity for both 28S and 18S rRNA after repair with RTCB (n=3 independent experiments). Hits were selected, when present in at least two replicates and when found by both analyses (<30 nt between 5′-end and 3′-end peak intensity). (I) Structural snapshots of the human ribosome (PDB 4ug0) . Shown in dark violet is the SRL as a reference point, the breakage sites identified by either analysis are shown in dark red, the space in between the mapped break sites is shown in light green. Error bars depict standard deviation (SD).

    Journal: bioRxiv

    Article Title: Eukaryotic tRNA ligases mediate RNA break repair

    doi: 10.64898/2026.05.26.727988

    Figure Lengend Snippet: (A) Left: Schematic of the direct RNA nanopore sequencing for the restrictocin cleaved and RTCB/Trl1 repaired ribosomes. Right: Schematic of sequence coverage plots of the 28S rRNA around the SRL cleavage site. After cleavage with restrictocin, the sequence coverage drops, this drop is reduced after break repair with both tRNA ligases. (B) The sequence coverage drop is reduced by repair with both tRNA ligases. The sequence coverage 5′ of the cleavage site (nt 4550) was normalized to the sequence coverage 3′ of the cleavage site (nt 4650) for all analyzed samples after nanopore sequencing. n=4 independent experiments, statistical significance of cleavage and repair was determined by One-way ANOVA followed by Tukey’s multiple comparison test. (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***). (C) Around 60% of cleaved HEK293T ribosomes were repaired by RTCB, whereas ∼70% of ribosomes were repaired by Trl1. (D) Schematic of the unspecific breakage and repair assay, using RNase 4, which cleaves between any accessible U/A, or U/G dinucleotide on the isolated ribosomes, followed by repair with RTCB or Trl1 and nanopore sequencing-based breakage and repair site-mapping. (E-H) Most RNase4-induced RNA breaks show reduced relative 5′-end peak intensity or 3′-end peak intensity for both 28S and 18S rRNA after repair with RTCB (n=3 independent experiments). Hits were selected, when present in at least two replicates and when found by both analyses (<30 nt between 5′-end and 3′-end peak intensity). (I) Structural snapshots of the human ribosome (PDB 4ug0) . Shown in dark violet is the SRL as a reference point, the breakage sites identified by either analysis are shown in dark red, the space in between the mapped break sites is shown in light green. Error bars depict standard deviation (SD).

    Article Snippet: Adherent HEK293T cells (CRL-3216, American Tissue Culture Collection, ATCC) (kind gift from the Christina Paulino lab) were grown at 37°C, 5% carbon dioxide and 70% humidity in 10cm dishes (Gibco) in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and antibiotic/antimycotic solution (Capricorn Scientific).

    Techniques: Nanopore Sequencing, Sequencing, Comparison, Isolation, Standard Deviation

    (A) Schematic of the experimental setup treating the RTCB knockdown cells with increasing concentrations of menadione for 3 h, before analyzing the isolated total cellular via capillary electrophoresis. (B) Electropherograms showing the effect of the RTCB knockdown with increasing concentrations of menadione. With increasing menadione concentrations, RNA is more fragmented in the RTCB KD compared to the WT. (C) Plots for the relative fluorescent units (a.u., arbitrary units) for three concentrations of menadione (0 µM, 60 µM and 100 µM) of the electropherograms shown in (B). Top: Whole cell RNA plot. Bottom: zoom in to small RNA fragments (nt 0-1500). (D) eCDF plots of cumulative probability for each read length after nanopore sequencing of RNA from cells treated with 0 µM and 60 µM menadione. Shown are read lengths for 28S rRNA (axis break between 2999 nt and 4500 nt) and 18S. (E) eCDF plots of cumulative probability for each read length after nanopore sequencing using specific sequencing adapters for RPPH1 and RN7SK RNA from cells treated with 0 µM and 60 µM menadione. (F) Puromycin incorporation assay for HEK293T cells, treated with harringtonine as a control, and 3 different concentrations of menadione (40 µM, 60 µM, 100 µM) for 1 h. Median fluorescence intensity (MFI) of puromycin-AF488 is shown. For gating strategy refer to Figure S5C. (G) Polysome profiling (15-50% sucrose gradient fractionation) using either untreated cells, or cells treated with 60 µM menadione for 1 h. For quantification, profiles are categorized in three sections: 1) free mRNPs and 40S subunits, 2) sub- and light polysomes and 3) heavy polysomes. The peaks for 40S and 60S subunits, as well as for 80S ribosomes and polysomes are indicated. (H) Quantification of the AUC of the polysome profiles of ribosome/total ratios according to the 3 categories. N=4 independent experiments. (I) Western blot analysis specific for RTCB, and the ribosomal proteins RPLS6 (40S) and RPL10A (60S) using sucrose fractions from (H). (J) Viability assay of RTCB WT and RTCB knockdown cells treated with increasing concentrations of menadione. With increasing concentrations of menadione, cells are less viable in the RTCB KD compared to the WT. n=7 biological replicates. (K) Area under the curve (AUC) for the viability plots in (F). The AUC for the viability in the KD cells in significantly lower compared to the WT. Statistical significances for all panels in this figure were determined using unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Journal: bioRxiv

    Article Title: Eukaryotic tRNA ligases mediate RNA break repair

    doi: 10.64898/2026.05.26.727988

    Figure Lengend Snippet: (A) Schematic of the experimental setup treating the RTCB knockdown cells with increasing concentrations of menadione for 3 h, before analyzing the isolated total cellular via capillary electrophoresis. (B) Electropherograms showing the effect of the RTCB knockdown with increasing concentrations of menadione. With increasing menadione concentrations, RNA is more fragmented in the RTCB KD compared to the WT. (C) Plots for the relative fluorescent units (a.u., arbitrary units) for three concentrations of menadione (0 µM, 60 µM and 100 µM) of the electropherograms shown in (B). Top: Whole cell RNA plot. Bottom: zoom in to small RNA fragments (nt 0-1500). (D) eCDF plots of cumulative probability for each read length after nanopore sequencing of RNA from cells treated with 0 µM and 60 µM menadione. Shown are read lengths for 28S rRNA (axis break between 2999 nt and 4500 nt) and 18S. (E) eCDF plots of cumulative probability for each read length after nanopore sequencing using specific sequencing adapters for RPPH1 and RN7SK RNA from cells treated with 0 µM and 60 µM menadione. (F) Puromycin incorporation assay for HEK293T cells, treated with harringtonine as a control, and 3 different concentrations of menadione (40 µM, 60 µM, 100 µM) for 1 h. Median fluorescence intensity (MFI) of puromycin-AF488 is shown. For gating strategy refer to Figure S5C. (G) Polysome profiling (15-50% sucrose gradient fractionation) using either untreated cells, or cells treated with 60 µM menadione for 1 h. For quantification, profiles are categorized in three sections: 1) free mRNPs and 40S subunits, 2) sub- and light polysomes and 3) heavy polysomes. The peaks for 40S and 60S subunits, as well as for 80S ribosomes and polysomes are indicated. (H) Quantification of the AUC of the polysome profiles of ribosome/total ratios according to the 3 categories. N=4 independent experiments. (I) Western blot analysis specific for RTCB, and the ribosomal proteins RPLS6 (40S) and RPL10A (60S) using sucrose fractions from (H). (J) Viability assay of RTCB WT and RTCB knockdown cells treated with increasing concentrations of menadione. With increasing concentrations of menadione, cells are less viable in the RTCB KD compared to the WT. n=7 biological replicates. (K) Area under the curve (AUC) for the viability plots in (F). The AUC for the viability in the KD cells in significantly lower compared to the WT. Statistical significances for all panels in this figure were determined using unpaired t-test (defined significance levels: p<0.05 = *, p<0.01 = **, p<0.001 = ***, p<0.0001= ****). Error bars depict standard deviation (SD).

    Article Snippet: Adherent HEK293T cells (CRL-3216, American Tissue Culture Collection, ATCC) (kind gift from the Christina Paulino lab) were grown at 37°C, 5% carbon dioxide and 70% humidity in 10cm dishes (Gibco) in Dulbecco’s modified Eagle’s medium (DMEM, Sigma-Aldrich), supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and antibiotic/antimycotic solution (Capricorn Scientific).

    Techniques: Knockdown, Isolation, Electrophoresis, Nanopore Sequencing, Sequencing, Control, Fluorescence, Fractionation, Western Blot, Viability Assay, Standard Deviation