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