Extended Data Fig. 2: Alteration of alternative splicing upon inhibition of polyamine synthesis is consistent across cell lines and rescued by polyamine supplementation.
From: Polyamine-dependent metabolic shielding regulates alternative splicing
a, b, LC-MS analysis of [¹³C]-methionine incorporation in spermidine and spermine (a), and semi-quantitative PCR analysis of alternative splicing events (b) in various cell lines treated with SAM486A (0.5 µM) for 48 h. c, d, Quantitative LC-MS analysis of polyamines (c), and z-scored (Abs(ΔPSI) > 10%, adj-p < 0.1) heat map of AS events (all events) (d) in DU145 shAMD1 cells treated with doxycycline (100 ng/mL, 2 days) and individually supplemented with putrescine, spermidine, or spermine (10 µM each) (Abs(ΔPSI) > 10 %, p < 0.1). e-g LC-MS metabolomics of total polyamines (putrescine, spermidine, spermine) (top) and methionine-derived [¹³C] incorporation (bottom) (e), semi-quantitative PCR gel of alternative splicing events (f), and densitometric quantification of long/short transcript ratios (g) showing the effects of SAM486A (1 µM, 48 h), DFMO (50 µM, 48 h), or their combination in DU145 cells. Abbreviations: Dox: doxycycline, ND: non-detectable, S: SAM486A; D: DFMO. Statistics: One-tailed student’s paired t-test was used for panel (a, c, e). One-tailed one-sample t-test was applied in panel (g). p: p-value. *p < 0.05, **p < 0.01, ***p < 0.001. Dollars represent a two-tailed paired t-test of the effect of SAM486A vs SAM486A + DFMO (g). $ p < 0.05, $$$ p < 0.01. Error bars: Mean ± S.E.M. n= independent biological replicates (a, c, e: 4; b: 3; f, g: 5).
