Global analysis by LC-MS/MS of N6-methyladenosine and inosine in mRNA reveal complex incidence

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FIGURE 2.
FIGURE 2.

ADAR1 and METTL 3 do not show cross-regulation in HEK293 and 293T cells. (A) Immunoblot analysis of protein expression in HEK293 cells treated with control (scrambled) or shRNA specific for ADAR1 mRNA. The levels of proteins were detected with specific antibodies listed in Materials and Methods. (B) Immunoblot analysis of protein expression in 293T cells transfected with control (scrambled) or METTL3 targeting siRNAs (METTL3 KD). Proteins were detected with specific antibodies listed in Material and Methods. (C) Levels of inosine, m6A, and m6Am in poly(A) RNA after KD of ADAR1 in HEK293 cell line (n = 3–4). (D) Levels of inosine, m6A, and m6Am in poly(A) RNA after KD of METTL3 in the 293T cell line (n = 4). Data are mean ± SD, unpaired two-tailed t-test. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001. The levels of nucleoside modifications were normalized to the total molar amount of canonical nucleosides N (N = C + U + G + A). Source data are available online for this figure.

This Article

  1. Published in Advance January 2, 2025, doi: 10.1261/rna.080324.124 RNA 31: 514-528

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