Extended Data Fig. 1: Additional data for Fig. 1.
ns: not significant, * p < 0.05, ** p < 0.01, *** p < 10−3, **** p < 10−4. Mann-Whitney U test, two sided. Data are presented as mean values ± SD. a) Queuine, preQ1, queuosine (Q) and N15-labeled q (queuineN15) MS calibration curves with linear fits of the log10 values. Additional calibration curves (used for group 2 samples) used the MS matrix optimized for plasma. The limit of detection is ~0.1 nM for queuine and preQ1. b) Short retrosynthetic chemical description of the different synthetic routes that led to the MS standard compounds used in this work. c) Deletion rate of PAQS-seq of Q-modifiable tRNAs of genus roseburia in the human stool sample with and without periodate (IO) treatment. d) Proliferation measurement by absorbance at 460 nm, HEK293T cells. Arrow indicates the time of addition of preQ1 and/or queuine. n = 8 biological replicates. e) Proliferation measurement by absorbance at 460 nm, HEK293T cells. Arrow indicates the time of addition of preQ1 and/or queuine. n = 8 biological replicates. f) Proliferation measurement by absorbance at 460 nm, HEK293T cells. The starting cells were mixtures of 0Q and 100Q cells. Arrow indicates the time of addition of preQ1. n = 8 biological replicates. g) Proliferation measurement by absorbance at 460 nm, MEF cells. Arrow indicates the time of addition of preQ1 and/or queuine. n = 8 biological replicates. h) Relative cell count of proliferation measurement, BMDC cells. Arrow indicates the time of addition of preQ1 and/or queuine. Cell counts are normalized to 0Q cells at t = 0. n = 5 replicates for each condition. i) Flow cytometry of Murine BMDCs. From left to right: the myeloid cell population is identified by forward and side scatter, doublets are excluded, and live cells are quantified by selecting the DAPI low population. n = 5 biological replicates. j) Normalized cell count or proliferation of mouse BMDC cells under indicated preQ1 and queuine concentrations. All data normalized to the average of 0,0 condition at day 1. n = 5 replicates for each condition. p values from left to right: 0.202, 6.09e-3, 6.09e-3. Source numerical data are available in.
