recombinant human il 1 α protein  (Bio-Techne corporation)

Journal: Frontiers in Endocrinology

Article Title: Syntaxin 4 protects islet β-cells from cytokine-induced senescence

doi: 10.3389/fendo.2026.1725252

Figure Lengend Snippet: STX4 protects against cytokine- and DNA damage-induced β-cell senescence. (A–D) Representative immunoblots (left panel) for STX4, phosphorylation of H2AX (γH2AX), p21 and tubulin (loading control). Right: Quantitation of STX4, γH2AX and p21 relative to tubulin. (A) Small interfering RNA knockdown of STX4 (siSTX4, for 72 h) in MIN6 cells. Control: siCtrl. (B) β-cell–specific adenoviral (AdRIP) overexpression of STX4 for 72 h followed by cytokine cocktail treatment (IL-1β, IFN-γ, and TNF-α) in MIN6. (C) STX4-overexpressing MIN6 cells (AdRIP-STX4) were exposed to bleomycin for 24 h (D) Human non-diabetic islets transduced with β-cell–specific STX4 and treated with a cytokine cocktail (48 h). (B–D) Control vector: AdRIP-Ctrl. Data represent mean ± SEM, each point corresponds to an independent biological replicate (n=4-5) * p <0.05, ** p <0.01, *** p <0.005, **** p <0.0001 using unpaired, two-tailed Student’s t-Test.

Article Snippet: Following transduction, islets were treated with a proinflammatory cytokine cocktail consisting of IL1-β (1 ng/mL, R&D System, cat #201-LB/CF, Minneapolis, MN), TNF-α (5 ng/mL, R&D System, cat #210-TA/CF), IFN-γ (40 ng/mL, R&D System, cat #285-IF/CF) for 48 h.

Techniques: Western Blot, Phospho-proteomics, Control, Quantitation Assay, Small Interfering RNA, Knockdown, Over Expression, Transduction, Plasmid Preparation, Two Tailed Test

Journal: Acta Pharmaceutica Sinica. B

Article Title: Inhibition of ASCT2 induces hepatic stellate cell senescence with modified proinflammatory secretome through an IL-1 α /NF- κ B feedback pathway to inhibit liver fibrosis

doi: 10.1016/j.apsb.2022.03.014

Figure Lengend Snippet: ASCT2 drives the proinflammatory SASP via IL-1 α /NF- κ B feedback loop through interacting with precursor IL-1 α at Lys82 of senescent HSCs. (A) Protein of p31–IL-1 α and p16–IL-1 α was measured by Western blot analysis with 4-OHT ( n = 3 per group). (B) Detection of intracellular IL-1 α by ELISA with 4-OHT ( n = 3 per group). (C) mRNAs of proinflammatory SASP including IL1A , IL1B , IL6 and IL8 were measured by qPCR ( n = 5 per group). (D) Co-IP of ASCT2 with IL-1 α in LX2 cells, as detected by Western blot analysis ( n = 3 per group). (E) Schematic of fragments of IL-1 α that were fused to GST (top) and were transfection into 293T cells. Binding of ASCT2 to GST-IL-1 α proteins was detected by Western blot (bottom, n = 3 per group). FL: full length; F1: domain 1–112; F2: domain 112–271; F3: NLS domain (80–88) deletion; F4: K82N mutation. (F) rIL-1 α protein was added to senescent (ASCT2 depletion or etoposide treatment) LX2 cells, and IL1R1 signaling cascade indicators were analyzed by Western blotting ( n = 3 per group). (G) rIL-1 α protein was added to senescent (ASCT2 depletion or etoposide treatment) LX2 cells, and NF- κ B protein was measured in nuclear and cytoplasmic lysates by Western blot ( n = 3 per group). (H) Relative luciferase measurement of NF- κ B in indicated cells ( n = 5–6 per group). (I) ASCT2 depletion-senescent LX2 cells were treated with rIL-1 α protein and addressed in NF- κ B inhibitor PDTC, and the analysis of mRNAs of proinflammatory SASP IL1A , IL1B , IL6 and IL8 by qPCR ( n = 5–6 per group). Bars indicate mean ± SEM; ∗ P < 0.05, ∗∗ P < 0.01; ns, no significance.

Article Snippet: Recombinant human IL-1 α protein (200-LA, protein for addition of IL-1 α ) was purchased from Bio-Techne (R&D Systems, Shanghai, China), the concentration used is 20 ng/mL in vitro .

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Transfection, Binding Assay, Mutagenesis, Luciferase