Journal: EMBO Molecular Medicine

Article Title: Tranilast directly targets NLRP 3 to treat inflammasome‐driven diseases

doi: 10.15252/emmm.201708689

Figure Lengend Snippet: Immunoblot analysis of ASC oligomerization in lysates of BMDMs treated with various doses (above lanes) of TR for 30 min and then stimulated with nigericin for another 30 min. Immunoprecipitation (IP) and immunoblot analysis of the interaction of endogenous NLRP3 and NEK7 in LPS‐primed BMDMs treated with various doses (above lanes) of TR for 30 min and then stimulated with nigericin for another 20 min. IP and immunoblot analysis of the interaction of endogenous NLRP3 and ASC in LPS‐primed BMDMs treated with various doses (above lanes) of TR for 30 min and then stimulated with nigericin for another 20 min. Data information: Data are representative of two or three independent experiments. Source data are available online for this figure.

Article Snippet: Recombinant human NLRP3 was from Novus Biologicals.

Techniques: Western Blot, Immunoprecipitation

Journal: EMBO Molecular Medicine

Article Title: Tranilast directly targets NLRP 3 to treat inflammasome‐driven diseases

doi: 10.15252/emmm.201708689

Figure Lengend Snippet: A, B Cell lysates of LPS‐primed BMDMs (A) or PMA‐differentiated THP‐1. (B) Cells were incubated with different concentrations of biotin‐TR for 1 h, which were then pulled down with streptavidin beads. C Purified recombinant NLRP3 protein was incubated with different concentrations of biotin‐TR for 1 h, which were then pulled down with streptavidin beads. D, E Cell lysates from HEK‐293T cells transfected with Flag‐tagged NLRP3, NOD1, NOD2, NLRP1, NLRC4, NLRP3‐LRR, NLPR3‐NACHT, or NLRP3‐PYD constructs were incubated with different concentrations of biotin‐TR for 1 h, which were then pulled down with streptavidin beads. F Immunoprecipitation (IP) and immunoblot analysis of the interaction of Flag‐NLRP3 and VSV‐NLRP3 in the lysates of HEK‐293T cells. TR was added at 8 h post‐transfection. G Immunoblot analysis of NLRP3 by SDD‐AGE or SDS–PAGE assay in BMDMs treated with TR for 30 min and then left stimulated with nigericin for 20 min. Data information: Data are representative of two or three independent experiments. Source data are available online for this figure.

Article Snippet: Recombinant human NLRP3 was from Novus Biologicals.

Techniques: Incubation, Purification, Recombinant, Transfection, Construct, Immunoprecipitation, Western Blot, SDS Page

Journal: EMBO Molecular Medicine

Article Title: Tranilast directly targets NLRP 3 to treat inflammasome‐driven diseases

doi: 10.15252/emmm.201708689

Figure Lengend Snippet: A, B FACS analysis of neutrophil numbers (A) or ELISA (B) of IL‐1β in the peritoneal cavity of 10‐week‐old male C57BL/6J mice intraperitoneally injected with MSU (1 mg/mouse) with or without TR (200 mg/kg of body weight). n = 8 per group. C, D Time course of changes of joint swelling (C) or ELISA (D) of IL‐1β in the supernatants of joint culture of 10‐week‐old male WT or Nlrp3 −/− mice intra‐articularly injected with MSU with or without oral TR administration (200 mg/kg of body weight). n = 6 per group. E Nlrp3A 350VneoR crossed with LysM‐Cre mice (NLRP3‐mut) treated with vehicle or TR at day 9. F Weight of wild‐type mice or Nlrp3A 350VneoR crossed with LysM‐Cre mice (NLRP3‐mut) treated with vehicle or TR at day 9. For WT vehicle and WT TR, n = 6; NLRP3‐mut PBS n = 4; and NLRP3‐mut TR n = 8. G Survival of NLRP3‐mut mice treated with vehicle or TR up to day 18. TR group n = 8, vehicle group n = 7. Data information: Data are shown as mean and s.e.m. and are representative of two or three independent experiments. Statistics were analyzed using an unpaired Student's t ‐test or a generalized Wilcoxon test (G): * P < 0.05, *** P < 0.001, NS, not significant.

Article Snippet: Recombinant human NLRP3 was from Novus Biologicals.

Techniques: Enzyme-linked Immunosorbent Assay, Injection

Journal: EMBO Molecular Medicine

Article Title: Tranilast directly targets NLRP 3 to treat inflammasome‐driven diseases

doi: 10.15252/emmm.201708689

Figure Lengend Snippet: A Body weights change of 6‐week‐old male wild‐type (WT) or Nlrp3 −/− mice that were first fed with HFD for 14 weeks and then treated with different dose of oral TR for 6 weeks. n = 6 per group. B, C Fed (B) or fasting (C) blood glucose concentrations at week 6 in the mice described in (A). n = 6. D–G ITT (D, F) or GTT (E, G) performed at week 6 in the mice described in (A). n = 6 per group. Data information: Data are shown as mean and s.e.m. and are representative of two independent experiments. Statistics were analyzed using an unpaired Student's t ‐test or two‐way ANOVA for (D–G): * P < 0.05, ** P < 0.01, *** P < 0.001, NS, not significant.

Article Snippet: Recombinant human NLRP3 was from Novus Biologicals.

Techniques:

Journal: Antioxidants

Article Title: Indapamide Increases IRS1 Expression and Modifies Adiponectin/NLRP3/PPARγ Crosstalk in Type 2 Diabetic Rats

doi: 10.3390/antiox11040691

Figure Lengend Snippet: Impact of daily CANA and INDA treatment on epididymal adipose tissue NLRP3 expression: oral CANA and INDA treatments were initiated once daily for 4 weeks. Data represent the mean ± S.E.M. (6 rats/group). ANOVA followed by post hoc Tukey–Kramer test at ( p < 0.05) was used for statistical comparison; a against normal CTRL, b against diabetic CTRL, d against diabetic/CANA.

Article Snippet: Moreover, using Cusabio ELISA kits (USA), NLRP3 (Cat. no. CSB-EL015871MO) and Nrf2 (Cat. no. CSB-E16188m) were quantified in the supernatant of the adipose tissue and the soleus muscle supernatant was used for the determination of IRS1 (Cat. no. CSB-PA047827) according to the enclosed pamphlet from the manufacturer.

Techniques: Expressing, Comparison