Extended Data Fig. 5: D-Cys decreases oxygen consumption rates and triggers ROS and lipid peroxidation in A549 cells.
From: d-cysteine impairs tumour growth by inhibiting cysteine desulfurase NFS1
a, b, Basal and maximal oxygen consumption rates (OCR) of BEAS-2B (a) or A549 cells (b) cultured in the absence (w/o) or presence of 500 μM D-Cys for three days. Data of n = 3 (a) and n = 4 (b) biological replicates are presented as mean ± SD and were analysed by 2-way ANOVA followed by Sidak’s multiple comparisons test. c, d, Quantitation of FACS-analyses of ROS production in A549 cells cultured in the absence (w/o) or presence of 500 μM D-Cys for three days, using either MitoSOX (determination of mitochondrial ROS in n = 3 biological replicates) (c) or BD C11 (determination of lipid peroxidation in n = 6 biological replicates) (d). Mean fluorescence intensity (MFI) ± SD is shown; data were analysed by unpaired two-tailed Student’s t-test. e, A549 cells have been cultured under normoxia (21% oxygen, O2) or hypoxia (1% oxygen, O2) for 72 h, either in the absence (w/o) or presence of 500 mM D-Cys. Cells were then counted (left panel) or stained with crystal violet (right panel). Cell counts of n = 3 biological replicates are presented as mean ± SD and were analysed by unpaired two-tailed Student’s t-test. f, Lack of effect of ferrostatin-1 (Fer-1) on D-Cys-induced toxicity. A549 cells were cultured for 72 h with or without D-Cys and with the indicated concentrations of Fer-1 added freshly from day 1 to day 3. Following the 72 h culture period cells of n = 3 biological replicates were counted, and the results are presented as mean ± SD. The comparison was performed by an unpaired two-tailed Student’s t-test.
