Article Title: LKB1 loss links serine metabolism to DNA methylation and tumourigenesis
Figure Lengend Snippet: Characterization of serine-glycine one-carbon pathway in KL cells a, Detailed graph of SGOC network. Enzymatic inhibitors used in this study are marked in red. b, Reactive oxygen species in K or KL cells transduced with shControl, shPSAT1-1 or shPSAT1-2 were measured by DCFDA (left) and CellRox staining (right). Data are normalized to cell number (n=4). c, Six-day proliferation assay of KL cells transduced with shControl, shPSAT1-1 or shPSAT1-2, showing lack of growth rescue by N-acetylcysteine (NAC). Data are expressed as relative to day 0 (n=6). d, Three-day growth assay of KL cells transduced with shControl, shPSAT1-1 or shPSAT1-2, showing the lack of rescue by excess nucleosides (adenosine, guanosine, thymidine, uridine, cytidine, 1 mM each). Data are presented as percentage of the growth of shControl cells (n=16). e, Proliferation of K or KL cells treated with aminooxyacetate (AOA). Data are expressed as relative to day 0 (n=8). f, Five-day proliferation of K or KL cells treated with AOA and/or NAC. Data are expressed as relative to day 0 (n=6). g, Data from RNA-seq ( left panel ) and quantitative proteomics ( right panel ) showing levels of genes involved in the production of SAM in K and KL. The data plotted are expressed as mean-centered values. h, Proliferation of K or KL cells treated with 2 mM cycloleucine. Data are expressed as percentage of the growth of vehicle treated cells (n=12). Shown are data pooled from two (b–f, h) experiments. For all panels, error bars are standard deviation unless otherwise stated and statistical significance is indicated as follows: *P
Article Snippet: Laminin (23017-95), 2NBDG (N13195), DCFDA (C6827), CellROX Green , CyQuant NF , bovine pituitary extract (13038-14), HBSS, RPMI, DMEM, DMEM:F12, MEM penicillin-streptomycin, pyruvate solution, fetal bovine serum (FBS) Alexa 488 and 568-conjugated secondary antibodies from Life Technologies; nicotinamide (N3376), trypsin inhibitor (T6522), dexamethasone (D4902), S-adenosylmethionine (A2408), 3,3,5-tri-iodo-L-thyronine (91990), oligomycin (75351), antimycin A (A8674), 2-deoxyglucose (D6134), dichloroacetate (347795), collagenase (C7657), glucose (G7528), aminooxyacetate , N-acetylcysteine (A9165), adenosine (A4036), guanosine (G6264), thymidine (T1895), uridine (U3003), cytidine (C4654), cycloleucine , cholera toxin (C8052), L-serine (S4500), betaine (61962), dimethylglycine (D1156), tamoxifen (T5148) and methylene blue (M4159) from Sigma; FCCP (0453), decitabine (2624), RG108 (3295), EGCG (4524), Compound C (3093), Torin 1 (4247) and galloflavin (4795) from Tocris; mouse EGF (354001), NuSerum IV (355104), ITS+ Premix (354352) and matrigel (354234) from Corning; SGI1027 (S7276) from Selleckchem; caspase-Glo 3/7 kit (G811A) and Celltiter-Glo kit (G755B) from Promega; 13 C-U-Glucose (CLM-1396-1) and 15 N-Glutamine (NLM-1016-1) from Cambridge Isotope Laboratories Inc; dispase (165-859) from Roche; lactate assay kit (K607-100) from Biovision Inc; Vectashield with DAPI (H1500) from Vector Laboratories; Bridge-IT SAM assay kit (1-1-1003B) from Medionics; meDIP kit (55009) from Active Motif; 3DZA (9000785) from Cayman Chemicals; Adeno-FlpO (1760) from Vector Biolabs; Adeno-Cre-eGFP from the Viral Vector Core Facility, University of Iowa.
Techniques: Transduction, Staining, Proliferation Assay, Growth Assay, RNA Sequencing Assay, Standard Deviation