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hek293 cells  (ATCC)


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    ATCC hek293 cells
    Hek293 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nacalai mtif3 ko hek293 cells
    Analysis of the effect of IF-3mt deletion on mitochondrial translation using <t>HEK293</t> wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).
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    Thermo Fisher hek293 cells
    IVT 5′-pppA RNAs stimulate the RIG-I/IFN pathway in human cells more efficiently than 5′-pppG RNAs. ( A ) To compare the RIG-I/IFN responses triggered by RNAs that begin with 5′-pppA and 5′-pppG, IVT RNA, representing a fragment of the IAV genome, was transfected into <t>HEK293</t> cells at various concentrations. The concentrations of type I IFN in the supernatants were assessed using the <t>HEK-Blue</t> IFN assay after 24 h. A selected concentration of RNA (100 ng/ml) was used to assess RIG-I levels and IRF3 phosphorylation with western blot upon treatment. ( C ) Dephosphorylated viral RNA (100 ng/ml) and Pol III transcript Y5 RNA were subjected to the HEK-Blue IFN assay. ( B , D , F ) Similar experiments were conducted in A549 cells, with type I IFN concentrations assessed in the supernatants and RIG-I levels and IRF3 phosphorylation in protein lysates in 8 h after treatment. The dotted line represents a lower quantification limit. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( A , B , E , F ) Upon log-transformation data were compared using two-way ANOVA with Šídák's multiple comparisons test. Experiments were conducted using either five ( A ), four ( E , F ) or three ( B ) biological replicates.
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    Jena Bioscience mtif3 ko hek293 cells
    Analysis of the effect of IF-3mt deletion on mitochondrial translation using <t>HEK293</t> wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).
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    Analysis of the effect of IF-3mt deletion on mitochondrial translation using HEK293 wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).

    Journal: Nucleic Acids Research

    Article Title: Selection of initiator tRNA and start codon by mammalian mitochondrial initiation factor 3 in leaderless mRNA translation

    doi: 10.1093/nar/gkaf021

    Figure Lengend Snippet: Analysis of the effect of IF-3mt deletion on mitochondrial translation using HEK293 wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).

    Article Snippet: HEK293 cells (ATCC, CRL-1573) and mtIF3 KO HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS), 1× sodium-pyruvate (Nacalai, 06977-34), and 0.05 g/l uridine (TCI, U0020) at 37°C with 5% CO 2 , according to the manufacturer’s instructions.

    Techniques: RNA Sequencing Assay, Synthesized, Two Tailed Test, Western Blot

    IVT 5′-pppA RNAs stimulate the RIG-I/IFN pathway in human cells more efficiently than 5′-pppG RNAs. ( A ) To compare the RIG-I/IFN responses triggered by RNAs that begin with 5′-pppA and 5′-pppG, IVT RNA, representing a fragment of the IAV genome, was transfected into HEK293 cells at various concentrations. The concentrations of type I IFN in the supernatants were assessed using the HEK-Blue IFN assay after 24 h. A selected concentration of RNA (100 ng/ml) was used to assess RIG-I levels and IRF3 phosphorylation with western blot upon treatment. ( C ) Dephosphorylated viral RNA (100 ng/ml) and Pol III transcript Y5 RNA were subjected to the HEK-Blue IFN assay. ( B , D , F ) Similar experiments were conducted in A549 cells, with type I IFN concentrations assessed in the supernatants and RIG-I levels and IRF3 phosphorylation in protein lysates in 8 h after treatment. The dotted line represents a lower quantification limit. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( A , B , E , F ) Upon log-transformation data were compared using two-way ANOVA with Šídák's multiple comparisons test. Experiments were conducted using either five ( A ), four ( E , F ) or three ( B ) biological replicates.

    Journal: Nucleic Acids Research

    Article Title: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs

    doi: 10.1093/nar/gkae1252

    Figure Lengend Snippet: IVT 5′-pppA RNAs stimulate the RIG-I/IFN pathway in human cells more efficiently than 5′-pppG RNAs. ( A ) To compare the RIG-I/IFN responses triggered by RNAs that begin with 5′-pppA and 5′-pppG, IVT RNA, representing a fragment of the IAV genome, was transfected into HEK293 cells at various concentrations. The concentrations of type I IFN in the supernatants were assessed using the HEK-Blue IFN assay after 24 h. A selected concentration of RNA (100 ng/ml) was used to assess RIG-I levels and IRF3 phosphorylation with western blot upon treatment. ( C ) Dephosphorylated viral RNA (100 ng/ml) and Pol III transcript Y5 RNA were subjected to the HEK-Blue IFN assay. ( B , D , F ) Similar experiments were conducted in A549 cells, with type I IFN concentrations assessed in the supernatants and RIG-I levels and IRF3 phosphorylation in protein lysates in 8 h after treatment. The dotted line represents a lower quantification limit. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( A , B , E , F ) Upon log-transformation data were compared using two-way ANOVA with Šídák's multiple comparisons test. Experiments were conducted using either five ( A ), four ( E , F ) or three ( B ) biological replicates.

    Article Snippet: HEK293 cells were transfected with short viral RNA and incubated for 24 h. After incubation total RNA was extracted with Trizol, DNA was removed using DNase I treatment and RNA was purified with GeneJET RNA Cleanup and Concentration Micro Kit (Thermo #K0841).

    Techniques: Transfection, Concentration Assay, Western Blot, Positive Control, Negative Control, Transformation Assay

    IVT 5′-pppA RNAs stimulate the RIG-I/IFN pathway in murine cells more efficiently than 5′-pppG RNAs. To compare the RIG-I/IFN responses triggered by RNAs that begin with 5′-pppA and 5′-pppG, IVT RNA, representing a fragment of the IAV genome, and Pol III transcript Y5 RNA were transfected into murine cells at a concentration of 100 ng/ml. ( A , B ) RIG-I levels and IRF3 phosphorylation in protein lysates were assessed using western blot. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. Upon log-transformation data were compared using two-way ANOVA with Šídák's multiple comparisons test. ( C , D ) The concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h after treatment ( n = 4). The dotted line represents a lower quantification limit. ( E ) Representative images of murine mKate+ BMDMs at 24 h upon RNA transfection. Scale bar is 100 μm. ( F ) Counts of mKate2+ cells at 0–48 h upon RNA transfection. Vertical lines represent standard deviation for n = 4. Longitudinal data upon shifted log-transformation (log 10 (x + 1)) were compared using repeated measures two-way ANOVA (rANOVA) with Geisser-Greenhouse correction and Šídák's multiple comparisons test.

    Journal: Nucleic Acids Research

    Article Title: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs

    doi: 10.1093/nar/gkae1252

    Figure Lengend Snippet: IVT 5′-pppA RNAs stimulate the RIG-I/IFN pathway in murine cells more efficiently than 5′-pppG RNAs. To compare the RIG-I/IFN responses triggered by RNAs that begin with 5′-pppA and 5′-pppG, IVT RNA, representing a fragment of the IAV genome, and Pol III transcript Y5 RNA were transfected into murine cells at a concentration of 100 ng/ml. ( A , B ) RIG-I levels and IRF3 phosphorylation in protein lysates were assessed using western blot. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. Upon log-transformation data were compared using two-way ANOVA with Šídák's multiple comparisons test. ( C , D ) The concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h after treatment ( n = 4). The dotted line represents a lower quantification limit. ( E ) Representative images of murine mKate+ BMDMs at 24 h upon RNA transfection. Scale bar is 100 μm. ( F ) Counts of mKate2+ cells at 0–48 h upon RNA transfection. Vertical lines represent standard deviation for n = 4. Longitudinal data upon shifted log-transformation (log 10 (x + 1)) were compared using repeated measures two-way ANOVA (rANOVA) with Geisser-Greenhouse correction and Šídák's multiple comparisons test.

    Article Snippet: HEK293 cells were transfected with short viral RNA and incubated for 24 h. After incubation total RNA was extracted with Trizol, DNA was removed using DNase I treatment and RNA was purified with GeneJET RNA Cleanup and Concentration Micro Kit (Thermo #K0841).

    Techniques: Transfection, Concentration Assay, Western Blot, Positive Control, Negative Control, Transformation Assay, Standard Deviation

    IVT-produced 5′-pppA RNAs contain more dsRNA compared with 5′-pppG RNAs. ( A ) Dot blot analysis of IVT short viral and Y5 RNAs was performed with anti-dsRNA J2 antibody, ( B ) then densitometric analysis ( n = 4 for short viral RNA, n = 3 for Y5 RNA) was conducted to assess the level of dsRNA byproducts in IVT-derived RNAs. ( C ) PAGE/Urea analysis was done on short viral RNA and Y5 RNA. ( D , C ) PAGE in native conditions was conducted on IVT control dsRNAs (produced by annealing of sense and antisense RNA strands) and ssRNAs. ( E ) Immuno-northern blot using PAGE native gel followed by transfer to the membrane and detection with anti-dsRNA J2 antibody was used to detect dsRNA. ( F ) PAGE in native conditions of IVT short viral RNAs and semisynthetic short viral RNAs produced by splint ligation revealed the presence of dsRNA in IVT 5′-pppA RNAs but not in semisynthetic counterparts. ( G ) To assess the immunogenic potential of splint-ligated RNAs, an IVT-produced antisense RNA, with or without triphosphate moieties, was added to the sense splint-ligated RNA. The concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h after treatment ( n = 3). The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( B , G ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test.

    Journal: Nucleic Acids Research

    Article Title: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs

    doi: 10.1093/nar/gkae1252

    Figure Lengend Snippet: IVT-produced 5′-pppA RNAs contain more dsRNA compared with 5′-pppG RNAs. ( A ) Dot blot analysis of IVT short viral and Y5 RNAs was performed with anti-dsRNA J2 antibody, ( B ) then densitometric analysis ( n = 4 for short viral RNA, n = 3 for Y5 RNA) was conducted to assess the level of dsRNA byproducts in IVT-derived RNAs. ( C ) PAGE/Urea analysis was done on short viral RNA and Y5 RNA. ( D , C ) PAGE in native conditions was conducted on IVT control dsRNAs (produced by annealing of sense and antisense RNA strands) and ssRNAs. ( E ) Immuno-northern blot using PAGE native gel followed by transfer to the membrane and detection with anti-dsRNA J2 antibody was used to detect dsRNA. ( F ) PAGE in native conditions of IVT short viral RNAs and semisynthetic short viral RNAs produced by splint ligation revealed the presence of dsRNA in IVT 5′-pppA RNAs but not in semisynthetic counterparts. ( G ) To assess the immunogenic potential of splint-ligated RNAs, an IVT-produced antisense RNA, with or without triphosphate moieties, was added to the sense splint-ligated RNA. The concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h after treatment ( n = 3). The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( B , G ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test.

    Article Snippet: HEK293 cells were transfected with short viral RNA and incubated for 24 h. After incubation total RNA was extracted with Trizol, DNA was removed using DNase I treatment and RNA was purified with GeneJET RNA Cleanup and Concentration Micro Kit (Thermo #K0841).

    Techniques: Produced, Dot Blot, Derivative Assay, Control, Northern Blot, Membrane, Ligation, Concentration Assay, Positive Control, Transfection, Negative Control

    Increased dsRNA production in 5′-pppA is independent of RNA structure and sequence. ( A ) Dot blot analysis of IVT produced variants of Y5 RNA starting from 5′-pppA or 5′-pppG was performed with anti-dsRNA J2 antibody, and ( B ) amount of dsRNA contamination was determined with densitometric analysis ( n = 3). ( C ) PAGE/Urea analysis of Y5 RNA variants showed single band for each RNA. ( D ) PAGE in native conditions, however, exhibited the presence of additional dsRNA bands. ( E ) Immuno-northern blot using PAGE native gel followed by transfer to the membrane and detection with anti-dsRNA J2 antibody was used to detect the presence of dsRNA formed by annealing with the 78 nt long complementary antisense strand. ( F ) HEK293 cells were transfected with Y5 RNA variants and the concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h n = 3). The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( B , F ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test.

    Journal: Nucleic Acids Research

    Article Title: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs

    doi: 10.1093/nar/gkae1252

    Figure Lengend Snippet: Increased dsRNA production in 5′-pppA is independent of RNA structure and sequence. ( A ) Dot blot analysis of IVT produced variants of Y5 RNA starting from 5′-pppA or 5′-pppG was performed with anti-dsRNA J2 antibody, and ( B ) amount of dsRNA contamination was determined with densitometric analysis ( n = 3). ( C ) PAGE/Urea analysis of Y5 RNA variants showed single band for each RNA. ( D ) PAGE in native conditions, however, exhibited the presence of additional dsRNA bands. ( E ) Immuno-northern blot using PAGE native gel followed by transfer to the membrane and detection with anti-dsRNA J2 antibody was used to detect the presence of dsRNA formed by annealing with the 78 nt long complementary antisense strand. ( F ) HEK293 cells were transfected with Y5 RNA variants and the concentration of type I IFN in the supernatants was measured using HEK-Blue IFN assay at 24 h n = 3). The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone. ( B , F ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test.

    Article Snippet: HEK293 cells were transfected with short viral RNA and incubated for 24 h. After incubation total RNA was extracted with Trizol, DNA was removed using DNase I treatment and RNA was purified with GeneJET RNA Cleanup and Concentration Micro Kit (Thermo #K0841).

    Techniques: Sequencing, Dot Blot, Produced, Northern Blot, Membrane, Transfection, Concentration Assay, Positive Control, Negative Control

    Long IVT-derived 5′-pppA RNAs contain more dsRNA byproducts compared with 5′-pppG RNAs. ( A ) Dot blot analysis of IVT EPO and Seg. 8 th of IAV PR8 RNAs starting from 5′-pppA or 5′-pppG and produced either with Class III (TATA) or Class II (TATT) T7 promoter. ( B ) Densitometric analysis ( n = 3 for EPO RNA, n = 4 for Seg. 8 th of IAV PR8 RNA) was conducted to assess the level of dsRNA byproducts in IVT-derived RNAs. ( C–F ) RIG-I/IFN pathway responses against EPO and Seg. 8 th of IAV PR8 RNAs starting from 5′-pppA or 5′-pppG were assessed with western blot analysis ( C , D ) and HEK-Blue IFN assay ( E , F ) 24 h after transfection into HEK293 cells ( n = 4). ( E , F ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test. The presence of dsRNA in EPO RNAs ( C , E , G ) RNAs ( D , F , H ). dsRNA controls are fully double stranded RNA derived from sense and antisense IVT. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone.

    Journal: Nucleic Acids Research

    Article Title: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs

    doi: 10.1093/nar/gkae1252

    Figure Lengend Snippet: Long IVT-derived 5′-pppA RNAs contain more dsRNA byproducts compared with 5′-pppG RNAs. ( A ) Dot blot analysis of IVT EPO and Seg. 8 th of IAV PR8 RNAs starting from 5′-pppA or 5′-pppG and produced either with Class III (TATA) or Class II (TATT) T7 promoter. ( B ) Densitometric analysis ( n = 3 for EPO RNA, n = 4 for Seg. 8 th of IAV PR8 RNA) was conducted to assess the level of dsRNA byproducts in IVT-derived RNAs. ( C–F ) RIG-I/IFN pathway responses against EPO and Seg. 8 th of IAV PR8 RNAs starting from 5′-pppA or 5′-pppG were assessed with western blot analysis ( C , D ) and HEK-Blue IFN assay ( E , F ) 24 h after transfection into HEK293 cells ( n = 4). ( E , F ) Data were compared using two-way ANOVA with Šídák's multiple comparisons test. The presence of dsRNA in EPO RNAs ( C , E , G ) RNAs ( D , F , H ). dsRNA controls are fully double stranded RNA derived from sense and antisense IVT. The positive control (C(+)) involved transfection with 100 ng/ml 3p-hpRNA, while the negative control (C(−)) was mock-transfected with lipofectamine alone.

    Article Snippet: HEK293 cells were transfected with short viral RNA and incubated for 24 h. After incubation total RNA was extracted with Trizol, DNA was removed using DNase I treatment and RNA was purified with GeneJET RNA Cleanup and Concentration Micro Kit (Thermo #K0841).

    Techniques: Derivative Assay, Dot Blot, Produced, Western Blot, Transfection, Positive Control, Negative Control

    Analysis of the effect of IF-3mt deletion on mitochondrial translation using HEK293 wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).

    Journal: Nucleic Acids Research

    Article Title: Selection of initiator tRNA and start codon by mammalian mitochondrial initiation factor 3 in leaderless mRNA translation

    doi: 10.1093/nar/gkaf021

    Figure Lengend Snippet: Analysis of the effect of IF-3mt deletion on mitochondrial translation using HEK293 wild-type cells (naïve) and IF-3mt KO cells. ( A ) The fold change of translation efficiency (ribosome footprint over RNA abundance) assessed by Ribo-seq and RNA-seq. The start codon of each ORF is indicated below. ( B ) Analysis of newly synthesized mitochondrial proteins by on-gel mito-FUNCAT. The signal intensities along the lanes are shown in the middle. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown on the right. The P- values were calculated by Student’s t -test (two-tailed). ( C ) Analysis of steady-state levels of selected mitochondrial proteins by western blotting. The start codon of each ORF is indicated below. TOM20, MRPS22/mS22, MRPL45, IF-2mt (nuclear encoded mitochondrial proteins), and α-Tubulin (cytoplasmic protein) were analyzed as references. The quantified relative amount of α-Tubulin is shown on the right. The means (bars), SDs (errors), and individual replicates ( n = 3, points) are shown. The P -values were calculated by Student’s t -test (two-tailed).

    Article Snippet: Naïve and mtIF3 KO HEK293 cells were cultured in six-well plates in methionine-free medium with 50 μM L-Homopropargylglycine (HPG) (Jena Bioscience) and 100 μg/ml anisomycin (Alomone Labs) for 3 h before cell harvest.

    Techniques: RNA Sequencing Assay, Synthesized, Two Tailed Test, Western Blot