mouse il 1β il 1f2 duoset elisa kit  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: MG53 protects against intestinal inflammation by inhibiting NLRP3 inflammasome activation

doi: 10.3389/fphar.2026.1791509

Figure Lengend Snippet: MG53 inhibits DSS-induced inflammation. (A) Representative images of immunofluorescence CD11b (red) and DAPI (blue) in WT and MG53 −/− mice on day 9 after DSS treatment. Scale bar, 50 μm. (B) Quantification of CD11b in panel (A) , n = 6 for each group. (C,D) Immunoblot (C) and quantification analysis (D) of mature interleukin-1β (IL-1β p17), interleukin-18 (IL-18) and cleaved caspase-1 (p20), in colon from WT and MG53 −/− mice on day 9 after DSS treatment. n = 6 each group. Data are shown as mean ± SD. Statistical significance was determined by unpaired two-tailed Student’s t-test. ** p < 0.01, * p < 0.05.

Article Snippet: The secretion of IL-1β from stimulated THP-1 cells or BMDMs was measured using human or mouse IL-1β ELISA kits (R&D Systems, DY201 for human, DY401 for mouse) following the manufacturer’s instructions.

Techniques: Immunofluorescence, Western Blot, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: MG53 protects against intestinal inflammation by inhibiting NLRP3 inflammasome activation

doi: 10.3389/fphar.2026.1791509

Figure Lengend Snippet: rhMG53 inhibits the activation of NLRP3 inflammasome. (A) Immunoblot of IL-1β p17 and caspase-1 p20 in supernatants (Sup.) from lipopolysaccharide (LPS)-primed mouse bone marrow–derived macrophages (BMDMs) (100 ng/mL, 3 h) treated with rhMG53 for 1 h and stimulated with nigericin for 1 h. Pro–IL-1β, pro–caspase-1, NOD-like receptor family pyrin domain containing 3 (NLRP3), and MG53 were analyzed in cell lysates. (B) IL-1β in culture supernatant was measured by enzyme-linked immunosorbent assay (ELISA). Treatments as in (A) . n = 3. (C) Immunoblot of IL-1β p17 and caspase-1 p20 in supernatants from LPS-primed BMDMs treated with rhMG53 (10 μg/mL, 3 h) and stimulated with nigericin (10 μg/mL, 1 h), adenosine 5′-triphosphate disodium salt (ATP) (5 mM, 30 min), or monosodium urate crystals (MSU) (150 μg/mL, 5 h). Pro–IL-1β, pro–caspase-1, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and MG53 were assessed in lysates. (D) ELISA of IL-1β release in culture supernatants. Treatments as in (C) . n = 3. (E) BMDMs were treated with rhMG53 (1, 5, 10 μg/mL) for 1 h before or after LPS stimulation, and NLRP3 inflammasome-related proteins were examined by immunoblotting. (F) Immunoblot of IL-1β p17 and caspase-1 p20 in culture supernatants of LPS-primed WT or MG53-overexpressing transgenic mice (TPA) BMDMs treated for 4 h and stimulated with nigericin (10 μg/mL, 1 h) or ATP (5 mM, 30 min). MG53, NLRP3, ASC, pro-IL-1β, and pro-caspase-1 were analyzed in cell lysates. (G) ELISA of IL-1β release in supernatants from LPS-primed WT or TPA BMDMs treated for 3 h and stimulated with nigericin (10 μg/mL, 1 h) or ATP (5 mM, 30 min). n = 5. (H) THP-1 cells differentiated with PMA were primed with LPS (0.5 μg/mL, 4 h), treated with rhMG53 at the indicated concentrations for 1 h, and stimulated with nigericin (1 μg/mL, 2 h). Supernatants and cell lysates were analyzed by immunoblotting. IL-1β p17 and caspase-1 p20 were detected in supernatants, while pro-caspase-1, pro-IL-1β, NLRP3, ASC, and MG53 were detected in lysates. (I) ELISA measurement of IL-1β release in culture supernatants. n = 3. Data are mean ± SD. Statistical significance was determined by one-way ANOVA (B,I) and unpaired Student’s t-test (D,G) , *** p < 0.005, ** p < 0.01, * p < 0.05.

Article Snippet: The secretion of IL-1β from stimulated THP-1 cells or BMDMs was measured using human or mouse IL-1β ELISA kits (R&D Systems, DY201 for human, DY401 for mouse) following the manufacturer’s instructions.

Techniques: Activation Assay, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transgenic Assay

Journal: Nature chemical biology

Article Title: A druggable redox switch on SHP1 controls macrophage inflammation

doi: 10.1038/s41589-026-02163-8

Figure Lengend Snippet: a, LPS-induced (100 ng/ml) SHP1 Tyr536, Tyr564 and Ser591 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. b , LPS-induced (100 ng/ml) STAT3 Tyr705 and Ser727 phosphorylation in iBMDMs over a time course of 2 h in iBMDMs pre-treated with DMSO or SCA1 (50 μM) for 3 h. Immunoblots shown are representative of three independent experiments. c,e,f , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs pre-incubated 3 h with SCA1, SCA1-NC or the respective derivatives over a dose response (0.313–80 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 4). For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, iBMDMs were pre-treated with LPS (100 ng/ml) for 3 h followed by treatment with DMSO or SCA9 at the indicated concentrations for 45 min and primed with adenosine triphosphate (ATP, 5 mM) for 45 min ( n = 3). Absolute cytokine production as a function of concentration is used to calculate IC 50 values. d , Pro-inflammatory cytokine IL-6 and TNF levels in cell supernatants of iBMDMs treated 3 h with SCA1 (20 μM) alone or 6 h with LPS (100 ng/ml) alone ( n = 3). g, Phagocytic activity of iBMDMs pre-treated 3 h with SCA1, SCA9, SCA7 or SCA25 (10 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 6). h, Relative fold changes in protein abundance between iBMDMs treated with SCA1 (50 μM) for 3 h followed by LPS (100 ng/ml, 15 min) versus LPS alone ( n = 3). Data are mean ± s.e.m. (in d,h ) or s.d. (in c,e-g ). P values calculated using one-way or two-way ANOVA for multiple comparisons or two-tailed Student’s t -tests for unpaired comparisons.

Article Snippet: For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, iBMDMs seeded and allowed to adhere as above are pre-treated with LPS (100 ng/ml) for 3 h followed by treatment with DMSO or SCA9 at the indicated concentrations for 45 min and primed with adenosine triphosphate (ATP, 5 mM) for 45 min. IL-1β levels in cell supernatants were quantified using mouse IL-1β DuoSet ELISA kit (R&D Systems, #DY401) according to manufacturer’s protocols.

Techniques: Phospho-proteomics, Western Blot, Incubation, Concentration Assay, Activity Assay, Quantitative Proteomics, Two Tailed Test

Journal: Nature chemical biology

Article Title: A druggable redox switch on SHP1 controls macrophage inflammation

doi: 10.1038/s41589-026-02163-8

Figure Lengend Snippet: a , Gating strategy for flow cytometry analysis of healthy donors or RA and MS patients PBMCs isolated monocytes using CD45, CD14 and CD16 staining ( n = 3 for each healthy and patient group). b , Percentage of CD45+CD14+CD16- monocyte population of interest ( n = 3 for each healthy and patient group). c , Pro-inflammatory cytokine TNF, IL-6 and IL-1β levels in cell supernatants of healthy donors and RA and MS patients monocytes pre-incubated 3 h with SCA1, SCA9, SCA7 and SCA25 (20 μM) followed by 6 h LPS stimulation (100 ng/ml) ( n = 3 for each healthy and patient group). For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, cells were additionally treated with adenosine triphosphate (ATP, 5 mM) for 45 min following LPS treatment. Data are mean ± s.e.m. P values calculated using two-tailed Student’s t -test for unpaired comparison or two-way ANOVA for multiple comparisons.

Article Snippet: For measurement of pro-inflammatory cytokine IL-1β levels in cell supernatants, iBMDMs seeded and allowed to adhere as above are pre-treated with LPS (100 ng/ml) for 3 h followed by treatment with DMSO or SCA9 at the indicated concentrations for 45 min and primed with adenosine triphosphate (ATP, 5 mM) for 45 min. IL-1β levels in cell supernatants were quantified using mouse IL-1β DuoSet ELISA kit (R&D Systems, #DY401) according to manufacturer’s protocols.

Techniques: Flow Cytometry, Isolation, Staining, Incubation, Two Tailed Test, Comparison