Journal: Molecular Cell

Article Title: METTL1 Promotes let-7 MicroRNA Processing via m7G Methylation

doi: 10.1016/j.molcel.2019.03.040

Figure Lengend Snippet:

Article Snippet: Recombinant METTL1/WDR4 (300 nM; Evotec) was incubated for 2 h with S-adenosylmethionine (15 μM) and oligonucleotide (1 μM) in a Tris-HCl pH 8.0 buffer (20 mM) supplemented with 1 mM DTT and 0.01% Triton X-100.

Techniques: Control, Virus, Recombinant, Protease Inhibitor, Gene Expression, Luciferase, Reverse Transcription, Labeling, RNA HS Assay, cDNA Synthesis, Knockdown, Expressing, Microarray, Imaging, Software, Extraction, Membrane, Autoradiography, Bradford Assay, Modification, Migration

Journal: Chemmedchem

Article Title: METTL3 Inhibitors for Epitranscriptomic Modulation of Cellular Processes

doi: 10.1002/cmdc.202100291

Figure Lengend Snippet: Protein thermal shift assay for METTL3/14 and METTL1 and cellular target engagement assay of UZH1a and UZH1b in HEK293T cells. (a) Results of the differential scanning fluorimetry (protein thermal shift assay) for METTL3/14 and METTL1 constructs. Derivative plot of the thermal denaturation curves of METTL3/14 and METTL1 in the presence of UZH1a ( red ), UZH1b ( gray ), or DMSO ( green ). Protein concentrations were 2 μM, mean±SD, N=3 (technical replicates). (b) Results of InCELL Pulse assay in HEK293T cells transfected with METTL3 construct and incubated with increasing concentrations of UZH1a and UZH1b for 1 h at 37 °C, followed by heating at 46 °C for 3 min. The soluble METTL3 fraction was quantified in luminescence‐based assay, DMSO‐treated cells served as normalization control, mean±SD, N=3–4 (biological replicates).

Article Snippet: The plasmid expressing the N‐terminally hexahistidine‐tagged METTL1 protein was a gift from Cheryl Arrowsmith (Addgene ID: 25264).

Techniques: Thermal Shift Assay, Drug discovery, Construct, Transfection, Incubation, Luminescence Assay, Control