Fig. 2: Sensitivity of MM cell lines to UNC1999 treatment was associated with metabolic alterations in methionine metabolism.
From: A distinct metabolic response characterizes sensitivity to EZH2 inhibition in multiple myeloma
a Effects of UNC1999 treatment on cell viability for the indicated cell lines. Biological replicates = 3. Statistical analysis was performed with one-way ANOVA. Values: mean with SEM. b Unsupervised clustering of seven human MM cell lines based on the abundance of all metabolites detected by LC-MS under basal conditions (DMSO treatment only). n(INA-6) = 9, n(U1996) = 11, n(KMS-28PE, U266-1970, LP-1, L363 and OPM2) = 4. c Representative western blot against H3K27me3 in the INA-6 (responsive) and U1996 (non-responsive) cells after UNC1999 treatment, and the relative signal quantification. d Western blot against H3 in the INA-6 (responsive) and U1996 (non-responsive) cells after UNC1999 treatment, and the relative signal quantification. The optical density in (d–e) was normalized against H4. Statistical analysis in (d–e) was performed with a two-tailed t-test. Values: mean with SEM. Data were collected from three biological replicates. The corresponding uncropped western blot figures (d–e) can be found in Supplementary Fig. 9c. e A principal component analysis (PCA) demonstrating in two dimensions that non-responding and responding samples cluster based on their independent metabolic profiles. n(DMSO INA-6) = 9, n(UNC1999 INA-6) = 9, n(DMSO U1996) = 11, n(UNC1999 U1996) = 10. f Heatmap showing the 150 most discriminating metabolites in the dataset, which were significantly different between INA-6 and U1996 after UNC1999 treatment. The colour bar describes the intensity of the change in metabolite levels: deeper colour determines the different magnitude of regulation. Red indicates upregulation and blue indicates downregulation. n(DMSO INA-6) = 9, n(UNC1999 INA-6) = 9, n(DMSO U1996) = 11, n(UNC1999 U1996) = 10. g–h Metabolite set enrichment analysis (MSEA) (g) in INA-6 and (h) non-responsive cell lines after UNC1999 treatment. n(DMSO INA-6) = 9, n(UNC1999 INA-6) = 9, n(DMSO KMS-28PE) = 4, n(UNC1999 KMS-28PE) = 4, n(DMSO U266-1970) = 4, n(UNC1999 U266-1970) = 4, n(DMSO u1996) = 11, n(UNC1999 U1996) = 10. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
