Fig. 4: SCD1-dependent viability for the mesenchymal M381 cells.
From: Raman-guided subcellular pharmaco-metabolomics for metastatic melanoma cells
a (Left) De novo synthesis pathway of monounsaturated fatty acids (MUFA) and (right) polyunsaturated fatty acids (PUFA) in mammalian cells. CAY10566 and SC 26196 are SCD1 (Δ9-desaturase) and Δ6-desaturase inhibitor, respectively. b Normalized (to 2908 cm−1) hSRS spectra of LDs in M381 cells without (CT) and with treatment of (top) 1, 5 and 10 μM CAY (n = 16, 18, 19, 19 for CT, 1, 5 and 10 μM CAY, respectively), and (bottom) 1 μM and 5 μM SC (n = 16, 19, 19 for CT, 1 μM and 5 μM SC, respectively) for 3 days. c Quantification of unsaturated lipid (UL) by intensity ratios of 3022 cm−1/2908 cm−1 from b. Relative viability of all five cell lines after treatment of 1 μM and 10 μM CAY for 3 days (n = 4 independent experiments) (d) or 1 μM and 10 μM SC for 3 days (n = 4 independent experiments) (e). f Relative viability of M381 cells after shRNA knockdown of SCD1 gene compared to scrambled control (CT) (n = 2 independent experiments). g GC-MS measurements of fatty acids extracted from bulk M381 cells with (CAY, purple) and without (CT, pink) treatment of 1 μM CAY for 3 days. The percentages of 16:0, 16:1, 18:0, 18:1, 18:2, and 20:4 fatty acids are normalized to total extracted fatty acids (n = 5 independent experiments). h Time-lapse apoptotic cell counts of M381 cells with (purple, CAY) and without (pink, CT) treatment of 1 μM CAY (n = 3 independent experiments, data shown as mean ± error with 95% CI). i Time-dependent relative viability of M381 cells after treatment of 1 μM CAY for 0, 1, 2, and 3 days (n = 4 independent experiments). j Normalized (to 2908 cm−1) hSRS spectra of LDs in M381 cells without (CT) and with 1 μM CAY treatment for 12 h, 1 day and 3 days (n = 20, 19, 17, 14 for CT, 12 h, 1 day and 3 days, respectively). k Quantification of UL from intensity ratios of 3022 cm−1/2908 cm−1 in j. **p < 0.01, ***p < 0.001, ns: not significant (p > 0.05) from two-tailed unpaired t-tests. Data shown as mean ± SEM. Source data are provided as a Source data file.
