Extended Data Fig. 9: Development of a bioavailable NNMTi.
From: NNMT inhibition in cancer-associated fibroblasts restores antitumour immunity
a, Human ovarian CAFs were treated with either NNMTi (5 µM) or DMSO in 5 µM L-methionine media, and 1-MNA levels were measured by mass spectrometry at the indicated time points (n = 3 technical replicates) (ordinary one-way ANOVA with Šídák’s multiple comparisons test). b, NNMT-expressing and Nnmt−/− control ovarian fibroblasts were treated for 72 h with NNMTi or DMSO. CAF marker expression (α-SMA, left) and cell size (FSC-A, right) were analysed by flow cytometry (n = 3 technical replicates) (ordinary one-way ANOVA with Šídák’s multiple comparisons test). MFI = mean fluorescence intensity. c-d, C57BL/6 mice (n = 3 per group) received a single treatment via oral gavage (p.o.) with varying concentrations of (c) NNMTi or (d) the less active distomer (NNMTi-D), and compound plasma levels were measured by mass spectrometry at the indicated time points. e-f, C57BL/6 mice (n = 3 per group) received a single p.o. treatment with varying concentrations of (e) NNMTi or (f) NNMTi-D, and 1-MNA plasma levels were measured by mass spectrometry. g, Human or murine NNMT, substrates, and assay reagents were mixed with different concentrations of NNMTi, and S-adenosylhomocysteine (SAH) levels were measured by MTase-GLO. Data are shown as mean ± SEM. One experiment representative of three independent experiments is shown for panel b.
