Journal: The Journal of Physiology

Article Title: Sensitization of colonic nociceptors by TNFα is dependent on TNFR1 expression and p38 MAPK activity

doi: 10.1113/JP283170

Figure Lengend Snippet: A , representative traces illustrating the concentration‐dependent increase in the magnitude of the normalized fluorescent response to TNFα (0.03–3.0 nM) within individual neurons from respective culture dishes. B , the mean magnitude of TNFα responses per dish increased across TNFα concentrations ( P = 0.050, one‐way ANOVA with Holm–Šídák's multiple comparisons test; n = 5 dishes from N = 5 independent cultures). C , the concentration‐dependent increase in the proportion (per dish) of TNFα‐responsive DRG neurons ( P = 0.014, the Kruskal–Wallis test with Dunn's multiple comparisons test; n = 5 dishes from N = 5 independent cultures), of which the majority were also co‐sensitive to capsaicin. D , the effects of TNFα (0.1 nM) were TNFR1‐mediated as illustrated by the significant reduction in ( E ) response magnitude ( P = 0.035; P = 0.012, one‐way ANOVA with Holm–Šídák's multiple comparisons test, n = 5−6 dishes from N = 5−6 independent cultures) and ( F ) proportion of TNFα‐responsive neurons following TNFR1 inhibition with 10 μM R7050 ( P = 0.004, one‐way ANOVA with Holm–Šídák's multiple comparisons test; n = 5−6 dishes from N = 5−6 independent cultures) or genetic deletion of TNFR1 in Tnfrsf1a –/– mice in comparison with neurons from wild‐type (WT) animals ( P = 0.0001, one‐way ANOVA with Holm–Šídák's multiple comparisons test; n = 5−6 dishes from N = 5−6 independent cultures). Dotted line represents the proportion of neurons activated in ECS controls (5.54 ± 5.01%, n = 6, N = 6). [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: R7050 (10 mM; DMSO), thapsigargin (1 mM; DMSO), A425619 (1 mM; DMSO) and SB203580 (10 mM; DMSO) were obtained from Tocris and stock concentrations made up as described.

Techniques: Concentration Assay, Inhibition, Comparison

Journal: Animals : an Open Access Journal from MDPI

Article Title: Staphylococcus aureus -Induced Necroptosis Promotes Mitochondrial Damage in Goat Endometrial Epithelial Cells

doi: 10.3390/ani12172218

Figure Lengend Snippet: The death receptor signaling and TLR4 did not contribute to the initiation stage of gEEC necroptosis. gEECs were pretreated with the TNFR1 inhibitor (R7050) and TLR4 inhibitor (TLR4-IN-C34), respectively, for 2 h, then infected with 10 MOI S. aureus for 4 h. ( A , B ) The cytotoxicity was determined using a CytoTox96 ® non-radioactive cytotoxicity assay kit. ( C ) Western blot analysis of P-MLKL. ( D ) Protein abundance of P-MLKL, normalized with β-actin. *** p < 0.001 vs. control group, # p < 0.05 vs. S. aureus group.

Article Snippet: For infection experiments, S. aureus was added to cells at a multiplicity of infection (MOI) of 10 or 100 for up to 6 h. For inhibition experiments, the cells were pretreated 1 h before infection with Necrostatin 2 recamate (Nec-1s, T7504) GSK’872 (T4074), Necrosulfonamide (NSA, T7129), TLR-IN-C34 (T8503), R7050 (T4637), Acetylcysteine (NAC, T0875), and rotenone (T2970) (TargetMol Biotech, Boston, MA, USA), glycine (1275, Biofroxx, Einhausen, Germany), and ionomycin (S1672, Beyotime Biotech, Beijing, China) at indicated concentrations.

Techniques: Infection, Cytotoxicity Assay, Western Blot