Fig 2: SF3B1 mutation promotes glycolytic metabolism in PDAC cells. (A) Gene set enrichment analysis based on Hallmark gene sets revealed gene sets related to SF3B1 mutation; false discovery rate was set at 0.25; NES indicates normalized enrichment score. (B) GSEA plot of MYC target and glycolysis related to SF3B1 mutation. (C‐E) Measurement of glucose uptake, lactate release, and ECAR in sh‐Ctrl and sh‐SF3B1 Panc05.04 cells (n = 3). (F) Effects of SF3B1‐WT or SF3B1‐K700E overexpression on the glucose uptake of Panc05.04‐sh‐SF3B1 cells (n = 3). (G) Effects of SF3B1‐WT or SF3B1‐K700E overexpression on the lactate release of Panc05.04‐sh‐SF3B1 cells (n = 3). Data were presented as the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001 by one‐way ANOVA and Tukey’s multiple comparisons test (C‐E) or Student’s t‐test (F, G).

Fig 3: Decreased c-Myc protein induced by matrine and rescued by proteasome inhibitor. a Effect of matrine on c-Myc protein expression in NK92 cells. NK92 cells were treated with matrine at 1.96 mM for 48 h, and c-Myc protein levels were measured by western blot. b c-Myc mRNA levels in NK92 cells were determined by quantitative RT-PCR at 24 h after matrine treatment at 0, 1.2, 2.4, and 3.6 mM (*p < 0.05, **p < 0.01 compared to 0 mM group). c CHX chase assay for the half-time of c-Myc. NK92 cells were treated with or without 1.96 mM matrine for 12 h. Cells were then treated with CHX (100 μg/mL) for the indicated minutes, and western blotting was performed. d c-Myc levels were quantified relative to GAPDH levels and graphed as percent c-Myc protein remaining after CHX treatment. Half-lives of c-Myc were calculated from exponential line equations and shown for each treat. e c-Myc protein levels were determined at 6 h post-treatment of MG132 and/or matrine, and (f) the relative intensities of target proteins were normalized to those of GAPDH. Analyses in triplicates. (*p < 0.05, **p < 0.01)

Fig 4: PRDX2 blocks apoptosis of myeloid leukemia cells. (A) Silenced PRDX2 using two independent shRNA and scrambled (shLuc) K562 cells were treated with 300 nM ABT-263 or without drug for 72 hours, and cell proliferation was measured by trypan blue exclusion method. (B) Colony formation ability was assayed in Methocult medium after being stably silenced of PRDX2 in K562 cells. After 7 days, colonies were photographed and counted manually; original magnification: ×100. (C) PRDX2-silenced and scrambled K562 cells treated with 300 nM ABT-263 for 72 hours. Apoptosis cells were analyzed using Annexin V and PI staining by flow cytometric analysis. Representative flow cytometry analysis (left panel) and the % of Annexin V and PI-positive cells are shown (right panel). (D) Immunoblot data showing the PRDX2 silenced efficiency in K562 cells and protein expression of P-MYC and MYC after knock down of PRDX2 in K562 cells. β-Actin was used as a control for equal loading. (E) Proliferation of U937 cells transduced with WT- or R882H and knocked down of PRDX2 from transduced cells. (F) Apoptosis analysis of R882H-expressed U937 cells knocked down with shLuc or shPRDX2 and treatment of 300 nM ABT-263 for 72 hours. (G) Immunoblot data showing the PRDX2 silenced efficiency in U937 cells transduced with R882H and protein expression of MYC and CDK1. β-Actin was used as a control for equal loading. All studies were repeated at least once. The mean ± S.D. of at least two replicates was plotted. *P < .05; **P < .03; ***P < .01.

Fig 5: Regulation of MYC and MYCBP expression is ALK-dependent in NSCLC cellsA. and B. Quantitative real-time PCR (qRT-PCR) analysis of MYC or MYCBP in NSCLC cells. NSCLC cell line panel includes ALK+ cell lines (H2228, H3122, STE-1), a ROS1 fusion line (HCC78), two KRAS cell lines (A549; G12S, H460; Q61H) and NRAS (H1299; codon 61), two mutant EGFR cell lines (H1650, HCC827; del 19). C. and D. Quantitative real-time PCR analysis of MYC or MYCBP in ALK+ cell lines in response to 24 hour treatment with 200nM TAE684. (**P < 0.01, *P < 0.05). E. Western blot analysis of H2228, H3122 and STE-1 cells after 24 hour treatment with 200nM TAE684. F. Western blot analysis of ALK+ cells treated for 24 hours with the indicated dose of crizotinib.