Journal: bioRxiv

Article Title: METTL2 forms a complex with the DALRD3 anticodon-domain binding protein to catalyze formation of 3-methylcytosine in specific arginine tRNA isoacceptors

doi: 10.1101/745240

Figure Lengend Snippet: DALRD3 forms a complex with METTL2A/B to bind distinct arginine tRNAs. (A) Immunoblot of Strep-DALRD3 purified from 293T cells expressed alone or in conjunction with FLAG-METTL2A/B. Immunoblot was probed with anti-TwinStrep and anti-FLAG antibodies. (B) Nucleic acid stain of RNAs extracted from the indicated input or purified samples after denaturing PAGE. The migration pattern of tRNAs, 5.8S rRNA, and 5S rRNA are denoted. (C) Northern blot analysis of the gel in (B) using the indicated probes. Input represents 2% of total extracts used for purification. The % enrichment represents the amount of RNA in the Strep purification that was recovered from the total input. The experiment was performed three times, with comparable results.

Article Snippet: RNA was then purified using RNA Clean and Concentrator Zymo-Spin IC Columns (Zymo Research). tRNA transcripts were visualized on a 15% polyacrylamide, 7M urea gel stained with SYBR Gold nucleic acid stain.

Techniques: Western Blot, Purification, Staining, Migration, Northern Blot