Figure 3.
(a) Schematic of human rRNA ac4C sites. (b) Sequencing analysis of ac4C-dependent mismatches in cDNAs generated from human rRNA helix 34 and (c) helix 45 following borohydride treatment and RT. Sequence corresponds to the cDNA sense strand. Vehicle = water; NaBH4 = 100 mM sodium borohydride; (+alkali) NaBH4 = alkali pretreatment (100 mM NaCO3, pH 10, 60 °C, 1 h), precipitation, then borohydride. (d) Relationship between misincorporation signal and stoichiometry of ac4C in rRNA and (e) a synthetic RNA harboring ac4C in an “ACA” sequence context. For all data, error bars indicate the standard deviation (n = 3). Primary data in the form of full sequencing traces is provided in the Supporting Information.