Journal: Molecular biology reports

Article Title: NIT2 overexpression predicts poor prognosis in tongue squamous cell carcinoma patients.

doi: 10.1007/s11033-019-05197-5

Figure Lengend Snippet: Fig. 1 Tissue microarray. a One hundred and three specimens were detected using tissue chip technique and 93 cases were finally avail- able. Two cases were adenocarcinoma, one case was lost during the experimental procedure, four cases exhibited serious defects, and three cases could not be tracked, as the complete follow-up data was not available. In the end, 93 samples were obtained from the two different tissue microarray blocks of TSCC. b NIT2 was highly expressed in tongue cancer tissues by immunohistochemical staining, mainly in cytoplasm

Article Snippet: The PVDF membrane was blocked for 1 h and then incubated with rabbit antibody against NIT2 (Proteintech, Chicago, IL, USA) or GAPDH antibody (Proteintech, Chicago, IL, USA) overnight at 4 °C.

Techniques: Microarray, Immunohistochemical staining, Staining

Journal: Molecular biology reports

Article Title: NIT2 overexpression predicts poor prognosis in tongue squamous cell carcinoma patients.

doi: 10.1007/s11033-019-05197-5

Figure Lengend Snippet: Fig. 3 Expression of NIT2 in TSCC by Western Blot and qPCR. N, adjacent normal tissues of tongue; T, TSCC tissue. (p = 0.026 (a), p < 0.001 (b))

Article Snippet: The PVDF membrane was blocked for 1 h and then incubated with rabbit antibody against NIT2 (Proteintech, Chicago, IL, USA) or GAPDH antibody (Proteintech, Chicago, IL, USA) overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Cancers

Article Title: Comprehensive Metabolic Profiling of MYC-Amplified Medulloblastoma Tumors Reveals Key Dependencies on Amino Acid, Tricarboxylic Acid and Hexosamine Pathways

doi: 10.3390/cancers14051311

Figure Lengend Snippet: D425MED orthotopic tumors preferentially use the glutaminase II (GTK) pathway over the glutaminase 1 (GLS1) pathway. ( A ) Cartoon illustrating how glutamine is metabolized through GLS1 to yield glutamate m + 6 isotope (green arrows). The glutaminase II pathway (blue arrows) uses glutamine transaminase K (GTK) to generate glutamate m + 1 by adding the amino group from labeled glutamine to alpha-KG. The Nit2 enzyme converts m + 6 alpha-ketoglutaramate (KGM) to m + 5 alpha-KG. This can in turn be converted to m + 5 glutamate by glutamate dehydrogenasae. ( B ) Bar graph showing glutamine-derived glutamate in D425MED cells in vitro, with predominance of m + 6. ( C ) Bar graph showing glutamine-derived glutamate in D425MED cells in flank tumors, showing increasing prominence of m + 1. ( D ) Bar graph showing glutamine-derived glutamate in D425MED cells in orthotopic tumors, showing predominance of m + 1 and near-absence of m + 6. ( E ) Bar graph comparing glutamine-derived glutamate in D425MED cells in orthotopic tumors and normal brain, showing predominance of m + 1 isotopologue. Abbreviations: normal (control) cerebellum = CTL CB; normal (control) cortex = CTL CTX; D425MED orthotopic tumor = D425MED brain; D425MED flank xenograft tumor = D425MED flank. The bar graph shows the mean intensities with the SD as error bar. Each group has three biological replicate samples. n.s. not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t -test.

Article Snippet: We used antibodies against GLUT1 (Novus biologicals (NB110-39113)), glutamine synthetase (Abcam (ab73593)); glutathione synthetase (abcam (ab91591); GTK (KAT1) (Santa Cruz Biotechnologies (sc-374531)), Nit2 (Origene (TA501138)) and beta actin (Santa Cruz (sc-47778)).

Techniques: Labeling, Derivative Assay, In Vitro, Control

Journal: Cancers

Article Title: Comprehensive Metabolic Profiling of MYC-Amplified Medulloblastoma Tumors Reveals Key Dependencies on Amino Acid, Tricarboxylic Acid and Hexosamine Pathways

doi: 10.3390/cancers14051311

Figure Lengend Snippet: GTK is the main enzyme responsible for glutathione synthesis in vivo. ( A – C ). Bar graphs showing the contribution of glutamine-derived carbons and nitrogens to glutathione in D425MED in different environments. The majority of glutathione was synthesized through GTK because the highest glutamine-derived glutathione was the m + 1 isotopologue, in all models. The predominance is most stark in orthotopic tumors, where the m + 6 isotopologue is virtually undetectable. ( D ) Bar graph showing increased glutamine-derived glutathione in orthotopic tumor compared to normal brain. ( E ) Bar graph showing m + 1 glutamate to glutamine ratio in D425MED cells in orthotopic tumors. ( F ) Western blot of glutaminase II pathway enzymes showing increased GTK (arrow) in D425MED orthotopic tumors (D425MED brain) compared to uninvolved cerebellum (CTL CB). The upper band is likely non-specific. ACTIN shows equal loading in all lanes. In contrast to GTK, we did not detect increased NIT2 or GLS1 protein expression MED tumor compared to normal cerebellum. Numbers above each blot show densitometry normalized to ACTIN. Abbreviations: normal (control) cerebellum = CTL CB; normal (control) cortex = CTL CTX; D425MED orthotopic tumor = D425MED brain; D425MED flank xenograft tumor = D425MED flank. The bar graphs show the mean intensities with the SD as error bar. Each group has three biological replicate samples. * p < 0.05, ** p < 0.01, Student’s t -test.

Article Snippet: We used antibodies against GLUT1 (Novus biologicals (NB110-39113)), glutamine synthetase (Abcam (ab73593)); glutathione synthetase (abcam (ab91591); GTK (KAT1) (Santa Cruz Biotechnologies (sc-374531)), Nit2 (Origene (TA501138)) and beta actin (Santa Cruz (sc-47778)).

Techniques: In Vivo, Derivative Assay, Synthesized, Western Blot, Expressing, Control