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( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), <t>NDUFV2</t> ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.
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( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), <t>NDUFV2</t> ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.
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( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), <t>NDUFV2</t> ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.
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( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), <t>NDUFV2</t> ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.
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( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), <t>NDUFV2</t> ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.
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Image Search Results


( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), NDUFV2 ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.

Journal: The Journal of Clinical Investigation

Article Title: METTL1-mediated m 7 G modification of valine tRNAs drives metabolic adaptation in pancreatic ductal adenocarcinoma cells

doi: 10.1172/JCI204312

Figure Lengend Snippet: ( A – E ) Distribution of ATP5H ( A ), NDUFC1 ( B ), NDUFV2 ( C ), SLC25A4 ( D ), and UCP2 ( E ) mRNAs across ribosome fractions (free RNA/RNP, 40S/60S/80S, light polysome, heavy polysome) from sgNC and sgMETTL1 BxPC-3 and MiaPaCa-2 cells, quantified by qRT-PCR. Bar graphs show the proportion of untranslated and translated fractions. ** P < 0.01 and *** P < 0.001, by 2-tailed, unpaired t test. ( F ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and METTL1-KO cells by Western blotting. ( G ) Relative protein levels of NDUFV2 and UCP2 in PDAC control and valine tRNA–KD cells by Western blotting. ( H and I ) Oxygen consumption rate in control and METTL1-KO groups from BxPC-3 ( H ) and MiaPaCa-2 ( I ) cells by Seahorse Mito Stress testing. *** P < 0.001, by 1-way ANOVA followed by Dunnett’s multiple-comparison test. ( J and K ) Oxygen consumption rate in control and valine tRNA–KD group from BxPC-3 ( J ) and MiaPaCa-2 ( K ) cells by Seahorse Mito Stress testing. *** P < 0.001, by1-way ANOVA followed by Dunnett’s multiple-comparison test. Data are representative of 3 independent experiments. All data are presented as the mean ± SD.

Article Snippet: The NDUFV2 expression vector (pcDNA3.1/NDUFV2/myc-His) was obtained from Addgene (127491).

Techniques: Quantitative RT-PCR, Control, Western Blot, Comparison