Journal: Nature Communications

Article Title: Ferroptotic damage promotes pancreatic tumorigenesis through a TMEM173/STING-dependent DNA sensor pathway

doi: 10.1038/s41467-020-20154-8

Figure Lengend Snippet: a Survival of Pdx1-Cre;Kras G12D/+ ;Gpx4 −/− (KCG) or Pdx1-Cre;Kras G12D/+ ; Gpx4 −/− ;Tmem173 −/− (KCGT) mice with or without control IgG or anti-8-OHG antibody treatment ( n = 10 mice/group; log-rank [Mantel–Cox] test). b Pancreatic weight of the indicated mice (6 months; n = 5 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). c Representative pancreas histology of the indicated mice. d Percentages of histological structures in the pancreas of the indicated mice ( n = 5 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). e Representative images of immunofluorescence staining of macrophages (red) in pancreas in indicated mice at the age of 3 months. f Relative gene expression in the pancreas of the indicated mice ( n = 3 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). g Percentage of abnormal telomeres by FISH analysis in ductal cells from KCG and KCGT mice at 3 months of age ( n = 5 mice/genotype). h mRNA expression of Tmem173 in indicated mice at 3 months of age with or without clophosome treatment ( n = 3 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). Data in b , d , and f – h are presented as mean ± SD. Data are from two or three independent experiments.

Article Snippet: To study the effects of 8-OHG inhibition on pancreatic tumorigenesis, 4–6 weeks old indicated that mice were randomly allocated into groups and injected i.p. with mouse monoclonal anti-8-OHG antibody (10 mg/kg; #GTX41980, RRID:AB_10732443, GeneTex) and control IgG2B (10 mg/kg; #MAB004, RRID:AB_357346, R&D Systems) per week for 12 weeks.

Techniques: Control, Immunofluorescence, Staining, Gene Expression, Expressing

Journal: Cell Reports

Article Title: Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity

doi: 10.1016/j.celrep.2018.08.006

Figure Lengend Snippet:

Article Snippet: The human/mouse Anti-UCP1 antibody (MAB6158, monoclonal Mouse IgG 2B Clone # 536435, R&D Systems) was conjugated to Alexa647 (ThermoFisher Alexa Fluor 647 antibody labeling kit A20186) and incubated with adipocytes at 1:300 for 1 h. Mouse IgG2B Alexa Fluor 647-conjugated Isotype Control antibody (R&D systems) was used as a control.

Techniques: Control, Recombinant, Blocking Assay, Antibody Labeling, Software, Flow Cytometry

Journal: Cell Death and Differentiation

Article Title: Next-generation hypomethylating agent SGI-110 primes acute myeloid leukemia cells to IAP antagonist by activating extrinsic and intrinsic apoptosis pathways

doi: 10.1038/s41418-019-0465-8

Figure Lengend Snippet: a–b Following 24 h of pretreatment with SGI-110 (MOLM-13: 50 nM, ML-2: 2 µM), AML cells were treated with ASTX660 (MOLM-13: 40 µM, ML-2: 5 µM) for 15 h. Death receptor expression on cell surface was determined by flow cytometric analysis after staining with PE-conjugated antibodies specific to each death receptor (open histograms) or with isotype-matched IgG controls (shaded gray histogram). Representative overlay histograms (a) and quantification of cell surface expression of death receptors (b) from three independent experiments performed in triplicate are shown. (c–h) ML-2 cells were transfected with non-targeting control siRNA (siCtrl) or siRNA against TNFR1, DR5, or FAS. (c–e) Cell surface expression of death receptors (open histograms) was analyzed by flow cytometry. Gray shaded histograms represent respective isotype controls. Representative overlay histograms are shown. (f–h) Following 24 h of pretreatment with 2 µM SGI-110, ML-2 cells were treated with 5 µM ASTX660 for 36 h. Treatments with 1 ng/ml TNFα and 5 µM ASTX660 (f), 10 ng/ml TRAIL (g) or 750 ng/ml hexameric FAS ligand (FASLG) (h) for 36 h were used as positive controls to demonstrate the efficacy of gene silencing. Cell death was determined by Annexin V-FITC staining and flow cytometry. In (b, f–h), mean and SD of three independent experiments performed in triplicate are shown. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cell surface expression of death receptors Flow cytometric analysis of cell surface expression of TNFR1, DR4, DR5, and FAS was performed using following phycoerythrin (PE)-conjugated antibodies: anti-TNFR1 (130-106-286, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-DR4 (FAB347P), anti-DR5 (FAB6311P, R & D Systems, Wiesbaden, Germany) and anti-FAS (556641, BD BioSciences, San Diego, CA, USA) and their respective isotype controls (IgG1 (IC002P), IgG2b (IC0041P, R & D systems, Wiesbaden, Germany)), REA control (130-113-462, Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Expressing, Staining, Transfection, Control, Flow Cytometry