Extended Data Fig. 3: METTL5/TRMT112-mediated 18 S rRNA m6A modification regulates 80 S ribosome assembly and global mRNA translation.
From: N6-methyladenosine (m6A) in 18S rRNA promotes fatty acid metabolism and oncogenic transformation
a, Western blot analysis of METTL5 and TRMT112 protein expression in HCC cells with or without METTL5 knockdown. b, Western blot analysis of METTL5 and TRMT112 protein expression in HCC cells with or without TRMT112 knockdown. c, Anti-m6A and anti-IgG IP-qPCR analysis of 18 S rRNA m6A modification of HepG2 cells with or without METTL5 knockdown. n = 3. (c-e,i) the unit for n is “replicates”. d,e, Anti-m6A and anti-IgG IP-qPCR analysis of 18 S rRNA m6A modification of HepG2 and Huh7 cells with or without TRMT112 depletion. n = 3. f, TAE agarose gel electrophoresis of RNAs from control and METTL5-knockdown HepG2 cells. g, Western blot analysis of puromycin incorporated into nascent peptides to monitor global mRNA translation in HepG2 cells with or without METTL5 depletion. Coomassie bright blue gel in the right panel serves as a control. h, Polysome profiling analysis showed that 80 S subunit peaks were significantly decreased upon METTL5 knockdown. i, Anti-RPL24 RIP-qPCR assay revealed that METTL5 knockdown significantly decreases the interaction between 18 S rRNA and RPL24. n = 3. In the histograms, the data are shown as the mean ± SD with P-values labeled on individual panels. P values were indicated by two-tailed unpaired Student’s t test for (c-e and i) in this figure.
