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. Author manuscript; available in PMC: 2022 Jul 12.
Published in final edited form as: Nat Cell Biol. 2021 Jul 12;23(7):684–691. doi: 10.1038/s41556-021-00709-7

Fig. 2 ∣. ALKBH7 catalyses m22G demethylation.

Fig. 2 ∣

a, Modification levels of m22G/A, m2G/A and m1G/A by LC–MS/MS for an RNA probe mimicking mt-Ile after in vitro treatment with recombinant human TRMT1. b, Modification levels of m22G/A in RNA before and after treatment with recombinant human ALKBH7. Left: mt-Ile ssRNA probe; **P = 0.0061 and **P = 0.0061 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. Right: folded mt-Ile tRNA probe; **P = 0.0034 and **P = 0.0035 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. c, Modification levels of m2G/A in RNA before and after treatment with recombinant human ALKBH7. Left: mt-Ile ssRNA probe; *P = 0.0148 and ***P = 0.0004 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. Right: folded mt-Ile tRNA probe; **P = 0.0016 and **P = 0.0018 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. Most m22G modifications were converted to G. d, Modification levels of m22G/A in a dsRNA probe mimicking the mt-Ile region in mt-dsRNA before and after treatment with recombinant human ALKBH7. ***P = 0.0002 and ***P = 0.0002 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. e, ALKBH7 mediates RNA m22G demethylation. f, Modification levels of m1A/A in a ssRNA probe mimicking the mt-Leu1 region in mt-dsRNA before and after treatment with recombinant human ALKBH7. ***P = 0.0005 and ***P = 0.0005 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. g, Modification levels of m1A/A in a dsRNA probe mimicking the mt-Leu1 region in mt-dsRNA before and after treatment with recombinant human ALKBH7. ***P = 0.0005 and ***P = 0.0008 for ALKBH7 5 μM and 10 μM versus input, respectively; unpaired, two-tailed t-test. For a–d,f,g, n = 3 biologically independent samples; data are presented as mean values ± s.d.