Fig. 7: Cytostatic effects by NAD+ treatment and chemotherapeutic agents.
From: NAD+ regulates nucleotide metabolism and genomic DNA replication
a,b, QIBC analysis of EdU-positive HeLa (a) and U2OS (b) cells treated for 24 h as indicated with 2 mM NAD+ in complete medium (FM) or minimal medium (LM) supplemented with glucose (Gluc) or glutamine (Gln). Cells quantified in a: FM, 2,055; FM + NAD+, 4,577; LM, 1,397; LM + NAD+, 1,500; Gluc, 2,803; Gluc + NAD+, 4,144; Gln, 1,367; Gln + NAD+, 2,086, cells quantified in b: FM, 1,678; FM + NAD+, 1,572; LM, 599; LM + NAD+, 649; Gluc, 808; Gluc + NAD+, 495; Gln, 737; Gln + NAD+, 1,479, from n = 2 (Gln) or 3 (all other conditions) biological replicates. Two-tailed Welch’s t-test. c, QIBC analysis of γH2AX foci, EdU intensity and DNA content in cells treated with control siRNA (CTRL) or si-SLC25A51 #1 and for 24 h with NAD+ as indicated. Representative experiment from two independent experiments. d, Relative cell number to cells treated only with control siRNA (si-CTRL) for each cell line. Cells were transfected with one of two si-SLC25A51 (#1 and #2) for 48 h and treated for 24 h with 2 mM NAD+. Mean + s.d., n = 4 biological replicates, two-tailed Student’s t-test. e, Relative cell number to non-treated (NT) cells. Cells were treated for 72 h with 1 µM PARPi and/or NAD+ as indicated. f, Relative cell number to non-treated (NT) cells. Cells were treated for 72 h with 80 µM NAD+, 1 µM 5-FU and/or gemcitabine (Gem) as indicated. g, Proposed model. Exogenous NAD+ initially boosts cellular metabolism and DNA replication, but after prolonged treatment, pyrimidine biosynthesis is impaired and pyrimidines are depleted. In e and f, mean + s.d., n = 6 biological replicates, two-tailed Welch’s t-test. For boxplots in a and b, centre line indicates the median; box limits indicate the 25th and 75th percentiles; minima and maxima of whiskers extend 1.5 times the inter-quartile range from 25th and 75th percentiles, respectively. Numerical data are available in source data files.
