Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Irisin inhibited the formation of NETs in STZ-induced T2DM mouse model and improved T2DM. A . Glucose tolerance test evaluated the glucose tolerance of T2DM mice treated with Irisin, N = 5; B . ELISA detected the plasma insulin levels of T2DM mice treated with Irisin, N = 5; C - D . ELISA detected the levels of Cit-H3 and MPO in the plasma of T2DM mice treated with Irisin, N = 5; E . ELISA detected the serum Irisin levels of T2DM mice treated with Irisin, N = 3; F - G . Western blotting detected the expression levels of related proteins in pyroptosis and phosphorylation levels of STING and IRE1a, N = 3; H . HE staining detected the morphology of pancreatic tissue in T2DM mice treated with Irisin, N = 3; I . Immunofluorescence staining detected the levels of Cit-H3 and MPO in pancreatic tissue of T2DM mice treated with Irisin, N = 3; ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Western Blot, Expressing, Phospho-proteomics, Staining, Immunofluorescence

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: NETs promoted high glucose-induced Min6 cell pyroptosis, but Irisin could inhibit NETs. A . ELISA detected insulin levels in Min6 culture media under different treatment conditions; B - D . Western blotting detected the expression levels of GSDMD-N and C-caspase-1 in Min6 cells under different treatment conditions; E - F . Flow cytometry detected Min6 cell pyroptosis under different treatment conditions; G . Sytox green staining marked the levels of NETs released under different conditions; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Flow Cytometry, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Pyroptosis induced by NETs was regulated by NLRP1. A . Western blotting was performed to detect the expression of related proteins in pyroptosis in Min6 cells with NLRP1 knocked down; B . ELISA was used to detect the LDH, IL-1βand IL-18; C - D . Flow cytometry was used to detect pyroptosis in NLRP1 knocked down Min6 cells induced by NETs; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Pyroptosis induced by NETs was regulated by STING/IRE1α. A . ELISA was used to detect the LDH, IL-1β and IL-18; B - C . Western blotting was performed to detect the expression of related proteins in pyroptosis in Min6 cells with C-176 added; D - E . Flow cytometry was used to detect pyroptosis in Min6 cells with C-176 added induced by NETs; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Pipersentan: A De Novo Synthetic Endothelin Receptor Antagonist that Inhibits Monocrotaline- and Hypoxia-Induced Pulmonary Hypertension

doi: 10.3389/fphar.2022.920222

Figure Lengend Snippet: Effects of Pipersentan on hPASMC Proliferation, Migration and Calcium Mobilization (A–D) hPASMCs were treated with DMSO, Pipersentan (0.01–100 μM), Macitentan (0.01–100 μM), SSP (0.2 μM) or Triton and exposed to DMSO or ET-1 (1 μM) for 24 h. (A,B) Proliferation was determined by detecting DNA synthesis in adherent cells ( n = 5). (C,D) Cytotoxicity was determined by detecting LDH released into the cell culture medium ( n = 5) (E,F) hPASMCs were treated with DMSO, Macitentan (10 μM) or Pipersentan (0.1–10 μM) and exposed to DMSO or ET-1 (1 μM) for 24 h. (E) Representative images of migrated cells (scale bars, 500 μm, Original magnification: 200). (F) Quantification of the crystal violet levels ( n = 3) (G) The antagonistic activity against hPASMCs was determined by detecting the intracellular calcium mobilization ( n = 3). The IC 50 of Macitentan and Pipersentan was 0.51 ± 0.56 nM and 1.66 ± 0.51 nM. The data are presented as the mean ± standard deviation, *** p < 0.001 vs. ET-1 (-); ### p < 0.001, ## p < 0.01, # p < 0.05 vs. ET-1 (+), n. s., not significant, ANOVA with Tukey’s post hoc in (A–F) ; n. s., not significant, unpaired Student’s t-tests in (G) ; SSP, staurosporine; Pip, Pipersentan; Mac, Macitentan; ET-1, endothelin-1; LDH, lactate dehydrogenase; DMSO, dimethyl sulfoxide.

Article Snippet: Cell proliferation and death levels were determined using the Cell Proliferation and Cytotoxicity Assay Kit (#CA1210, Solarbio, China) and Cytotoxicity Lactate Dehydrogenase Detection Kit (#BC0685, Solarbio, China), respectively, according to the kit’s protocols.

Techniques: Migration, DNA Synthesis, Cell Culture, Activity Assay, Standard Deviation

Journal: Acta Neuropathologica Communications

Article Title: Activity of translation regulator eukaryotic elongation factor-2 kinase is increased in Parkinson disease brain and its inhibition reduces alpha synuclein toxicity

doi: 10.1186/s40478-018-0554-9

Figure Lengend Snippet: Effects of eEF2K inhibition on human AS cytotoxicity in differentiated N2A cells. a - b Western blot analysis of p-eEF2 (T56) levels in N2A cells subsequent to transient overexpression of human wild type or mutant A53T AS, with or without siRNA mediated eEF2K knockdown ( a ), and corresponding densitometry analysis ( b ) ( n = 6–9/group from three independent experiments; One-way ANOVA post-hoc Bonferroni test, * p < 0.05, *** p < 0.005; error bars indicate Mean ± S.E.M). c Measurements of cytotoxicity by lactate dehydrogenase-LDH release in the culture medium ( c ) and FACS analysis of propidium iodide-PI staining ( d ) in N2A cells subsequent to transient overexpression of human wild type or mutant A53T AS, with or without siRNA mediated eEF2K knockdown ( n = 9–12/group from three independent experiments; One-way ANOVA post-hoc Bonferroni test, * p < 0.05, ** p < 0.01, *** p < 0.005, NS = not significant; error bars indicate Mean ± S.D.)

Article Snippet: Additional reagents and biochemical assays employed during these studies include: pool of small interference RNAs (SiRNAs) targeting mouse eEF2K (Santa Cruz, #sc-39,012), Cell Titer Glo ATP measurement kit (Promega, #G7570), Lactate dehydrogenase (LDH) fluorometric assay (Novus Biologicals, #NBP2–54851), Seahorse Mito stress test kit (Agilent, #103015–100), 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) fluorescent ROS reagent (ThermoFisher, #D399), and MitoTracker Green fluorescent reagent for mitochondrial mass (ThermoFisher, #M7514).

Techniques: Inhibition, Western Blot, Over Expression, Mutagenesis, Knockdown, Staining