Journal: Hepatology Communications

Article Title: Proteomics‐based identification of the role of osteosarcoma amplified‐9 in hepatocellular carcinoma recurrence

doi: 10.1002/hep4.1952

Figure Lengend Snippet: Differential proteins expression in OS‐9 overexpression group (n = 3) and vector control (n = 3) of SMMC‐7721 cells. (A) Volcano plot of the differentially expressed proteins. Gray dots represent genes that are not differentially expressed in the early recurrence group and nonrecurrence group; red dots and blue dots represent genes that are up‐regulated and down‐regulated significantly in the early recurrence group. (B) Heat map of the differentially expressed proteins. Red rectangles mean that genes are up‐regulated in these samples, and blue ones mean down‐regulated. Two hundred and sixty‐eight protein expressions were up‐regulated and 140 protein expressions were down‐regulated in the overexpression group compared with the vector control group (fold change ≥ 1.5; p < 0.05). (C) Heat map of the differentially expressed proteins classified to the hypoxia‐inducible factor 1 (HIF‐1) and tumor necrosis factor (TNF) signaling pathway. (D) Ridgeline plot of Kyoto Encyclopedia of Genes and Genomes pathway enrichment for differentially expressed proteins. (E) Gene Ontology functions for differentially expressed proteins. The left side of the circle includes all related genes, and the right side displays the Gene Ontology terms. Red and blue rectangles mean that genes are up‐regulated and down‐regulated in the early recurrence group. Abbreviations: ALDOA, aldolase, fructose‐bisphosphate A; ALDOC, aldolase, fructose‐bisphosphate C; BCL10, BCL10 immune signaling adaptor; CASP7, caspase 7; CCNB1, cyclin B1; CDK6, cyclin dependent kinase 6; CEBPB, CCAAT enhancer binding protein beta; CHEK2, checkpoint kinase 2; CREBBP, CREB binding protein; DDX58, DExD/H‐box helicase 58; ENO1, enolase 1; ENO2, enolase 2; ENO3, enolase 3; HK2, hexokinase 2; HMOX1, heme oxygenase 1; IGFBP3, insulin like growth factor binding protein 3; JUNB, JunB proto‐oncogene; LDHA, lactate dehydrogenase A; MALT1, MALT1 paracaspase; MLKL, mixed lineage kinase domain like pseudokinase; OE, overexpressed; PGK1, phosphoglycerate kinase 1; PLCG2, phospholipase C gamma 2; SERPINE1, serpin family E member 1; SFN, stratifin. NC, control; STEAP3, STEAP3 metalloreductase; TNFAIP3, TNF alpha induced protein 3; TRAF5, TNF receptor associated factor 5

Article Snippet: Inhibitors including HIF‐1α and tumor necrosis factor alpha (TNF‐α) were obtained (VH‐298, MedChemExpress; HY‐100947, Methylthiouracil; HY‐B0513, MedChemExpress).

Techniques: Expressing, Over Expression, Plasmid Preparation, Control, Binding Assay

Journal: Hepatology Communications

Article Title: Proteomics‐based identification of the role of osteosarcoma amplified‐9 in hepatocellular carcinoma recurrence

doi: 10.1002/hep4.1952

Figure Lengend Snippet: (A–K) The relative messenger RNA (mRNA) expression level of lineage kinase domain‐like MLKL (A), tumor necrosis factor alpha–induced protein 3 (TNFAIP3) (B), JunB proto‐oncogene (JUNB) (C), and tumor necrosis factor receptor–associated factor 5 (TRAF5) (D), TNF‐α (E) and caspase‐7 (CASP7) (F) could be observed to increase more than 2‐fold ( p < 0.01). The relative expression of enolase1 (ENO1) (G), enolase2 (ENO2) (H), enolase3 (ENO3) (I), aldolase, fructose‐bisphosphate A (ALDOA) (J), and lactate dehydrogenase A (LDHA) (K) were also elevated more than 2‐fold ( p < 0.01)

Article Snippet: Inhibitors including HIF‐1α and tumor necrosis factor alpha (TNF‐α) were obtained (VH‐298, MedChemExpress; HY‐100947, Methylthiouracil; HY‐B0513, MedChemExpress).

Techniques: Expressing

Journal: The Journal of Experimental Medicine

Article Title: NLRP3 signaling drives macrophage-induced adaptive immune suppression in pancreatic carcinoma

doi: 10.1084/jem.20161707

Figure Lengend Snippet: NLRP3 expression in human and mouse PDA. (A) Lysate from 3-mo-old WT, KC, and KC;NLRP3 −/− mice were tested for expression of IL-1β and IL-18 by Western blotting. Ponceau staining is shown. Experiments were repeated three times. Representative data are shown. (B) Frozen sections of pancreata of mouse PDA tumors were tested for coexpression of CD11b and NLRP3 or CK19 and NLRP3 compared with respective isotype controls. Bar, 10 µm. (C) F4/80 + Gr1 − CD11c − CD11b + macrophages from pancreata or spleen macrophages from 3-mo-old KC mice were tested for expression of NLRP3 compared with isotype controls. (D) Macrophages from pancreata or spleen of KC mice were tested for coexpression of MHC II and CD206. (E) MHC II − CD206 + and MHC II + CD206 − pancreatic macrophage subsets from 3-mo-old KC mice were gated and tested for expression of NLRP3 and IL-1β. Representative and quantitative data are shown. Positive gates are based on isotype controls (not depicted). (F) MHC II − CD206 + and MHC II + CD206 − TAM subsets from WT mice bearing orthotopic PDA were gated and tested for expression of NLRP3 and IL-1β. (G) Macrophages from WT control pancreata or pancreata or spleen of WT mice harboring orthotopic KPC tumors were tested for expression of NLRP3. (H) Paraffin-embedded sections of human PDA were tested for expression of NLRP3 compared with isotype control. Bar, 20 µm. (I) CD15 + monocytic cells from single-cell suspensions of human PDA or PBMCs were gated by flow cytometry and tested for expression of NLRP3. Representative contour plots and quantitative data from six patients are shown. (J) Splenic macrophages from WT mice were cultured alone or in a 5:1 ratio with KPC-derived tumor cells. At 24 h, macrophages were tested for expression of CD206, IL-10, and NLRP3. (K) Similarly, BMDMs from WT mice were stimulated with recombinant TGF-β or TNF and tested for NLRP3 expression. (L) Orthotopic PDA-bearing mice were serially treated with a neutralizing TGF-β mAb or isotype control. Tumors were harvested on day 21, and expression of NLRP3 and CD206 in TAMs was determined by flow cytometry. n = 5/group. All mouse experiments were repeated a minimum of twice using five mice per experimental group. Littermate controls were used. Unpaired Student’s t test was used for statistical analyses. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Data are presented as mean ± standard error. MΦ, macrophage; Ms, mouse; Panc, pancreas; PanIN, pancreatic intraepithelial neoplasia; SSA, side scatter.

Article Snippet: In some experiments, day-10 BMDMs were treated with 8 pg/ml of recombinant mouse TNF (Cell Signaling Technology) or 0.2 ng/ml TGF-β (R&D Systems) for 18 h before analysis for NLRP3 expression by flow cytometry.

Techniques: Expressing, Western Blot, Staining, Control, Flow Cytometry, Cell Culture, Derivative Assay, Recombinant