Additional file 1 of ASS1 and ASL suppress growth in clear cell renal cell carcinoma via altered nitrogen metabolism

Additional file 1: Figure S1. ASS1 and ASL expression affects survival in ccRCC patients. A. Survival curve based on ASS1 expression in patients from the TCGA. (High n=176, low n=352). B. Survival curve based on ASL expression in patients from the TCGA (High n=258, low n=270). C. Log2 scale fold change in ASS1 and ASL protein levels in tumors when compared to normal kidney (n=108, Clark et al.)

Additional file 2 of ASS1 and ASL suppress growth in clear cell renal cell carcinoma via altered nitrogen metabolism

Additional file 2: Figure S2. ASL Expression Suppresses Growth in a ccRCC cells. A. Ectopic expression of ASL in 786-O polyclonal population. Actin is used as the loading control. B. 3D soft agar colony forming assay with 786-O cells expressing empty vector control and ASL cDNA. **p< 0.01. C. Ectopic expression of ASL in UMRC2 polyclonal population. Actin is used as the loading control. D. 3D soft agar colony forming assay with UMRC2 cells expressing empty vector control and ASL cDNA. **p< 0.01

Additional file 4 of ASS1 and ASL suppress growth in clear cell renal cell carcinoma via altered nitrogen metabolism

Additional file 4: Figure S4. Re-expression of ASL Alone in 786-O Cells Does Not Suppress Growth in vivo. A. 786-O subcutaneous xenograft tumor growth in nude mice, where cells expression ASL or empty vector. (n = 5). B. Immunohistochemistry staining and quantification for phosphorylated histone H3 as marker for proliferation in xenograft tumors. Scale bars are at 100μm. C. Immunohistochemistry staining and quantification for cleaved caspase 3 as marker for apoptosis in xenograft tumors. Scale bars are at 100μm. D. Immunohistochemistry staining and quantification for p21 as marker for proliferation in xenograft tumors. Scale bars are at 100μm. E. Immunohistochemistry staining and quantification for CD31 as marker for vasculature in xenograft tumors. Scale bars are at 100μm. *p< 0.05. F. Heat map showing the levels of various angiogenic proteins in 786-O xenograft tumors expressing ASL or empty vector

Additional file 3 of ASS1 and ASL suppress growth in clear cell renal cell carcinoma via altered nitrogen metabolism

Additional file 3: Figure S3. ASL Expression in 769-P cells Alters Amino Acid Pools and Affects DNA Replication. A. Steady-state amino acid levels in 769-P cells expressing empty vector and ASL cDNA. Error bars represent SEM of 3 technical replicates. ****p< 0.001, ***p< 0.001. B. Representative images of cell cycle analyses of 769-P cells with propidium iodide (PI) staining indicating a sub-G0 peak in cells expressing ASL cells at 0 hours and 24 hours. C. Exogenously provided pyrimidines (cytidine + thymidine, 50 μM) partially rescue growth in 769-P cells expressing ASL. Error bars represent SEM of 7 wells. *p< 0.05, ***p< 0.001. D. 786-O cells expressing ASS1+ASL cDNA were cultured with Trolox (Trol, 100μM), N-Acetylcysteine (NAC, 1mM), and/or nucleosides (adenosine + guanosine + cytidine + thymidine, 50μM) and assessed for cell proliferation by WST-1. Error bars represent SEM of 7wells. ***p< 0.001

DS_10.1177_0363546518778789 – Supplemental material for Modulating Glucose Metabolism and Lactate Synthesis in Injured Mouse Tendons: Treatment With Dichloroacetate, a Lactate Synthesis Inhibitor, Improves Tendon Healing

<p>Supplemental material, DS_10.1177_0363546518778789 for Modulating Glucose Metabolism and Lactate Synthesis in Injured Mouse Tendons: Treatment With Dichloroacetate, a Lactate Synthesis Inhibitor, Improves Tendon Healing by Kairui Zhang, Michael W. Hast, Soutarou Izumi, Yu Usami, Snehal Shetye, Ngozi Akabudike, Nancy J. Philp, Masahiro Iwamoto, Itzhak Nissim, Louis J. Soslowsky and Motomi Enomoto-Iwamoto in The American Journal of Sports Medicine</p

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Supplementary Data_v1 from Metabolic Enzyme DLST Promotes Tumor Aggression and Reveals a Vulnerability to OXPHOS Inhibition in High-Risk Neuroblastoma

Supplemental Method and Materials; Supplemental Figure Legends; Goes with the Longer version of the paper</p