Journal: PloS one

Article Title: Pepsin digest of wheat gliadin fraction increases production of IL-1β via TLR4/MyD88/TRIF/MAPK/NF-κB signaling pathway and an NLRP3 inflammasome activation.

doi: 10.1371/journal.pone.0062426

Figure Lengend Snippet: Figure 1. PDWGF digest induces IL-1b, IL-18, and IL-1a release in monocytes and PBMC from CD patients. IL-1b (A), IL-18 (B) and IL-1a (C) levels were quantified in cell supernatants by ELISA. Data are given as mean 6 SD from 39 patients and 15 healthy donors (HD). *P,0.05, **P,0.01 (CD vs. HD). (D). PDWGF-induced activation of PBMC is not due to LPS contamination. Results are shown as the percentage of the cytokine production from 4 CD patients. The data were normalized to the result from untreated cells which was set as 100%. Mean 6 SD, n = 4 independent experiments, ***P,0.001 compared to untreated cells. doi:10.1371/journal.pone.0062426.g001

Article Snippet: The concentrations of human IL-1b, IL-1a, IL-18 and TNF-a as well as murine IL-1b and TNF-a were measured by commercial ELISA Duo Set Kits (R&D Systems) or ELISA MAX kits (Biolegend) according to manufacturer instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Activation Assay

Journal: PloS one

Article Title: Pepsin digest of wheat gliadin fraction increases production of IL-1β via TLR4/MyD88/TRIF/MAPK/NF-κB signaling pathway and an NLRP3 inflammasome activation.

doi: 10.1371/journal.pone.0062426

Figure Lengend Snippet: Figure 3. PDWGF-induced IL-1b production from celiac patient PBMC is modulated by K+ efflux, but is independent of the P2X7 receptor; as shown by (A) ELISA, mean 6 SD, n = 10, ***P,0.001 vs. PDWGF-treated cells; and by (B) Western blot. Representative blots from 5 independent experiments are shown. (C) Inhibition of ROS modulate PDWGF-induced IL-1b secretion, mean 6 SD, n = 10; as well as (D) pro-IL-1b production from PBMC of CD patients. Representative blots from 3 independent experiments are shown. b-actin was used as a loading control. ***P,0.001 vs. PDWGF- treated cells. doi:10.1371/journal.pone.0062426.g003

Article Snippet: The concentrations of human IL-1b, IL-1a, IL-18 and TNF-a as well as murine IL-1b and TNF-a were measured by commercial ELISA Duo Set Kits (R&D Systems) or ELISA MAX kits (Biolegend) according to manufacturer instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Inhibition, Control

Journal: PloS one

Article Title: Pepsin digest of wheat gliadin fraction increases production of IL-1β via TLR4/MyD88/TRIF/MAPK/NF-κB signaling pathway and an NLRP3 inflammasome activation.

doi: 10.1371/journal.pone.0062426

Figure Lengend Snippet: Figure 4. MAPK and NF-kB are involved in PDWGF mediated IL- 1b secretion. PDWGF-primed PBMC were treated with SB203580 (SB), SP600125 (SP), UO125 (UO), TPCK, or PDWGF, alone or in combination for 24 h. (A) IL-1b was quantified in cell supernatants by ELISA. Mean 6 SD, n = 5 independent experiments. ***P,0.001 vs. PDWGF-treated cells. (B) Pro-IL-1b levels were examined in cell lysates by immunoblot- ting from 5 experiments. b-actin was used as a loading control. doi:10.1371/journal.pone.0062426.g004

Article Snippet: The concentrations of human IL-1b, IL-1a, IL-18 and TNF-a as well as murine IL-1b and TNF-a were measured by commercial ELISA Duo Set Kits (R&D Systems) or ELISA MAX kits (Biolegend) according to manufacturer instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: American Journal of Physiology - Renal Physiology

Article Title: ELABELA antagonizes intrarenal renin-angiotensin system to lower blood pressure and protects against renal injury

doi: 10.1152/ajprenal.00606.2019

Figure Lengend Snippet: ELA-32 treatment inhibits the activation of intrarenal renin-angiotensin system in high salt (HS)-loaded Dahl salt-sensitive (SS) rats. A: representative immunoblotting and densitometric analysis of renal (pro)renin receptor (PRR) and cyclooxygenase-2 (COX-2) expression in different experimental groups; expression was normalized by β-actin. B: quantitative RT-PCR analysis of PRR, renin, AGT, ACE, AT1R, AT2R, and ACE2 mRNA expression in the kidney cortical and medullary; expression was normalized by GAPDH. C: urinary and plasma soluble PRR (sPRR) levels. D: urinary and plasma prorenin/renin levels. E: urinary and plasma angiotensinogen (AGT) levels. F: urinary and plasma angiotensin II (ANG II) levels. Male Dahl SS rats were used in these studies. n = 5 Rats per group. Data are means ± SE. *P < 0.05, **P < 0.01, ***P < 0.001 vs. normal Na+ (NS); #P < 0.05, ##P < 0.01, ###P < 0.001 vs. HS. Statistical analysis was performed by using 1-way analysis of variance with the Bonferroni test using IBM SPSS 19 software. ELA, ELABELA; fPRR, full-length PRR.

Article Snippet: To detect urinary, plasma, and cell culture medium soluble PRR (sPRR), total renin/prorenin, and cAMP, urinary and plasma AGT and ANG II, urinary ELA, apelin, PGE 2 , neutrophil gelatinase-associated lipocalin (NGAL), and albumin, we used the commercially available enzyme immunoassay (EIA) kits according to the manufacturers’ instructions: soluble (Pro) renin Receptor Assay Kit (cat. no. 27782; Immuno-Biological, Gunma, Japan), Rat Prorenin and Renin Total Antigen ELISA Kit (cat. no. RPRENKT-TOT; Molecular Innovations), Cyclic AMP ELISA Kit (cat. no. 581001; Cayman Chemical), Rat Angiotensinogen ELISA Kit (cat. no. MBS762162; MyBioSource), ANG II ELISA kit (cat. no. ADI-900-204; Enzo Life Sciences, Farmingdale, NY), [pGlu 1 ]-ELA-32 (Human) EIA Kit (cat. no. EK-007-19; Phoenix Pharmaceuticals), Mouse Apelin EIA Kit (cat. no. {"type":"entrez-protein","attrs":{"text":"Q9R0R4","term_id":"20137544","term_text":"Q9R0R4"}} Q9R0R4 ; RayBiotech Life), Prostaglandin E 2 ELISA Kit (cat. no. 514010; Cayman Chemical), and Rat NGAL ELISA Kit (cat. no. 501530; Cayman Chemical), respectively.

Techniques: Activation Assay, Western Blot, Expressing, Quantitative RT-PCR, Software