Extended Data Fig. 9: Cell viability in combined treatments.
From: NAD+ regulates nucleotide metabolism and genomic DNA replication
a, Immunoblot detection of SLC25A51 depletion in U2OS and HeLa cells. Cells were treated with control siRNA (si-CTRL) or one of two si-SLC25A51 (#1 and #2). M denotes lane with molecular weight marker. b, c, Immunoblot confirmation of mitochondrial (mito.) isolation from HeLa (b) and U2OS (c) cells treated for 24 h with control siRNA (si-CTRL) or SLC25A51-targeting siRNA and with 2 mM NAD+ as indicated. d, Mitochondrial NAD(H) quantification in cells treated as in b, c, NT = non-treated. Data are mean+SD, n = 3 (HeLa) and n = 2 (U2OS) biological replicates, two-tailed Student’s t-test (HeLa). e, Relative cell number of HeLa and U2OS cells treated for 24 h without (NT) or with brequinar (BRQ) as indicated. Data are mean+SD, n = 6 biological replicates, two-tailed Welch’s t-test. f, Relative cell number of U2OS cells treated for 72 h without (NT) or with 1 µM PARPi and/or NAD+ as indicated. Data are mean+SD, n = 6 biological replicates, two-tailed Welch’s t-test. g, Relative cell number of HeLa cells treated for 72 h without (NT) or with 2 mM NAD+ and/or 10 µM PARPi. Data are mean+SD, n = 3 biological replicates, two-tailed Student’s t-test. h-j, Percentage dead cells for HeLa (h), MRC5 (i) and U2OS (j) cells treated as in f. Data are mean+SD, n = 6 biological replicates, two-tailed Welch’s t-test. k, Relative cell number of U2OS cells treated for 72 h without (NT) or with 80 µM NAD+, 1 µM 5-fluorouracil (5-FU) and/or gemcitabine (Gem) as indicated. Data are mean+SD, n = 6 biological replicates, two-tailed Welch’s t-test. l-n, Percentage dead cells for HeLa (l), MRC5 (m) and U2OS (n) cells treated as in k. Data are mean+SD, n = 6 biological replicates, two-tailed Welch’s t-test. Immunoblots were performed twice with similar results. Numerical data and uncropped immunoblots are available in Source Data Files.
