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Mettl3

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METTL3 is a protein that functions as a methyltransferase enzyme, catalyzing the addition of methyl groups to RNA molecules. It plays a role in post-transcriptional modifications of RNA.

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Lab products found in correlation

Sybr green, by Thermo Fisher Scientific (2 mentions) Map3k6, by Novus Biologicals (2 mentions) Mapk14, by Cell Signaling Technology (2 mentions) α actinin antibody, by Merck Group (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Map4k5, by Thermo Fisher Scientific (2 mentions) Gapdh, by Biosynth (2 mentions) Fitc conjugated wheat germ agglutinin, by Merck Group (1 mentions) Fluorescein isothiocyanate conjugated wheat germ agglutinin, by Merck Group (1 mentions) G3bp1, by Santa Cruz Biotechnology (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Trans blot turbo transfer system, by Bio-Rad (1 mentions)

Close spelling variants of this product

Anti-METTL3

3 protocols using mettl3

1

Western Blotting and Immunostaining of Neonatal Rat Cardiomyocytes

Cited 1 time
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Western blotting was performed from neonatal rat cardiomyocytes using standard procedures. Antibodies used were METTL3 (Bethyl Laboratories), MAP3K6 (Novus Biologicals), MAP4K5 (Thermo Scientific Pierce), MAPK14 (Cell Signaling Technology) and GAPDH (Fitzgerald Industries). Masson’s trichrome staining was performed from histological sections generated from paraffin-embedded hearts. Immunostaining was performed using α-Actinin antibodies (Sigma Aldrich) and cell area was quantified using CellProfiler following published methods.24 (link) Detection of the cell membrane was performed using FITC-conjugated Wheat Germ Agglutinin (Sigma Aldrich). Cross-sectional area was measured using ImageJ. RNA was extracted from neonatal rat cardiomyocytes or mouse hearts using Trizol and reverse transcription was performed using the High Capacity cDNA Reverse Transcription kit (Applied Biosystems). Selected genes and m6A peaks were analyzed by real-time PCR using SYBR green (Applied Biosystems). Quantified mRNA expression was normalized to Rpl7 (Ribosomal Protein L7), and expressed relative to controls.
Dorn L.E., Lasman L., Chen J., Xu X., Hund T.J., Medvedovic M., Hanna J.H., van Berlo J.H, & Accornero F. (2018). The N6-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy. Circulation, 139(4), 533-545.
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2

Cardiac Protein and Gene Expression Analysis

Cited 1 time
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Western blotting was performed from neonatal rat cardiomyocytes using standard procedures. Antibodies used were METTL3 (Bethyl Laboratories), MAP3K6 (Novus Biologicals), MAP4K5 (Thermo Scientific Pierce), MAPK14 (Cell Signaling Technology), and GAPDH (Fitzgerald Industries). Masson trichrome staining was performed from histological sections generated from paraffin-embedded hearts. Immunostaining was performed using α-actinin antibodies (Sigma-Aldrich), and cell area was quantified using CellProfiler following published methods.24 (link) Detection of the cell membrane was performed using fluorescein isothiocyanate–conjugated wheat germ agglutinin (Sigma-Aldrich). The cross-sectional area was measured using ImageJ. RNA was extracted from neonatal rat cardiomyocytes or mouse hearts using Trizol, and reverse transcription was performed using the High Capacity cDNA Reverse Transcription kit (Applied Biosystems). Selected genes and m6A peaks were analyzed by real-time polymerase chain reaction using SYBR green (Applied Biosystems). Quantified mRNA expression was normalized to Rpl7 (ribosomal protein L7) and expressed relative to controls.
Dorn L.E., Lasman L., Chen J., Xu X., Hund T.J., Medvedovic M., Hanna J.H., van Berlo J.H, & Accornero F. (2018). The N6-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy. Circulation, 139(4), 533-545.
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3

Protein Transfer and Western Blot Analysis

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SDS-PAGE gels were blotted onto nitrocellulose membrane with the Trans-Blot Turbo transfer system (Bio-Rad). Blots were stained with Ponceau stain to confirm equal loading of proteins. Blots were blocked in 5% skim milk in PBS. Primary antibodies to the following proteins were used: G3BP1 (Santa Cruz Biotechnology; sc-98561), Flag (Sigma-Aldrich; F1804), GST (Thermo; MA4-004) and METTL3 (Bethyl Laboratories; A301-567-A). Images were processed in ImageJ. Western blot quantifications were done with ImageJ.
Edupuganti R.R., Geiger S., Lindeboom R.G., Shi H., Hsu P.J., Lu Z., Wang S.Y., Baltissen M.P., Jansen P.W., Rossa M., Müller M., Stunnenberg H.G., He C., Carell T, & Vermeulen M. (2017). N6-methyladenosine (m6A) recruits and repels proteins to regulate mRNA homeostasis. Nature structural & molecular biology, 24(10), 870-878.
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