nf-κb inhibitor sn50  (AnaSpec)

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Liver Sinusoidal Endothelial Cells Promote Metabolic Dysfunction-associated Steatohepatitis Progression via Interleukin-1R1-mediated Chemokine Production Induced by Macrophage-derived Interleukin-1β

doi: 10.1016/j.jcmgh.2025.101698

Figure Lengend Snippet: Increased IL-1β secretion by macrophages due to autophagy impairment enhances inflammatory chemokine expression in LSECs. ( A ) Protein expression in ATG7-KO macrophages and NC macrophages generated using the CRISPR-CAS9 system. ( B ) Evaluation of IL-1β and caspase-1 levels in the culture supernatant of THP-1 macrophages by ELISA following 24-hour stimulation with or without 25 ng/mL LPS (n = 3/group). ( C ) Experimental method using LPS-stimulated THP-1 macrophage conditioned medium. ( D ) Evaluation of NF-κB or MAP kinase-related proteins in TMNK-1 cells. ( E and F ) Inflammatory chemokine gene expression in TMNK-1 cells when treated with control medium or THP-1 macrophage conditioned medium (n = 4/group). SN50, NF-κB inhibitor; SP600125, JNK inhibitor; SB203580, p38 MAPK inhibitor. ( G ) Inflammatory chemokine gene expression measured 24 hours after treating TMNK-1 cells with varying concentrations of human recombinant IL-1β (n = 4/group). ( H ) Evaluation of inflammatory chemokine expression in TMNK-1 cells after 6 hours of stimulation with recombinant human IL-1β (5 ng/mL) in the presence of SP600125 and SB203580 (n = 4/group).

Article Snippet: The inhibitors used were as follows: the NF-κB inhibitor SN50 (HY-P0151; MedChemExpress), the JNK inhibitor SP600125 (HY-12041; MedChemExpress), the p38 MAPK inhibitor SB 203580 (HY-10256; MedChemExpress), and the IL-1R antagonist (IL-1Ra) (093-05991; FUJIFILM Wako Pure Chemical Corporation).

Techniques: Expressing, Generated, CRISPR, Enzyme-linked Immunosorbent Assay, Gene Expression, Control, Recombinant

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Liver Sinusoidal Endothelial Cells Promote Metabolic Dysfunction-associated Steatohepatitis Progression via Interleukin-1R1-mediated Chemokine Production Induced by Macrophage-derived Interleukin-1β

doi: 10.1016/j.jcmgh.2025.101698

Figure Lengend Snippet: Macrophage-derived IL-1β increases CCL2 expression in HSCs via JNK activation. ( A ) Evaluation of the levels of NF-κB or MAPK-related proteins in LX-2 cells. ( B, C ) CCL2 gene expression in LX-2 cells treated with the control medium or THP-1 macrophage conditioned medium (n = 3/group). SN50, NF-κB inhibitor; SP600125, JNK inhibitor; SB203580, p38 MAPK inhibitor. ( D ) Evaluation of CCL2 expression in TMNK-1 cells after 48 hours of stimulation with recombinant human IL-1β (5 ng/mL) in the presence of SP600125 and SB203580 (n = 4/group).

Article Snippet: The inhibitors used were as follows: the NF-κB inhibitor SN50 (HY-P0151; MedChemExpress), the JNK inhibitor SP600125 (HY-12041; MedChemExpress), the p38 MAPK inhibitor SB 203580 (HY-10256; MedChemExpress), and the IL-1R antagonist (IL-1Ra) (093-05991; FUJIFILM Wako Pure Chemical Corporation).

Techniques: Derivative Assay, Expressing, Activation Assay, Gene Expression, Control, Recombinant

Journal: CytoJournal

Article Title: 1α, 25-dihydroxyvitamin D3 attenuates tumor necrosis factor-α-induced endothelial cell injury by modulating the tumor necrosis factor-α/nuclear factor kappa-B pathway

doi: 10.25259/Cytojournal_220_2024

Figure Lengend Snippet: Effects of 1α,25-(OH)2D3 on the NF-κB signaling pathway. (a) Effect of NS50 on the expression of TNF-α-induced cell adhesion molecules (Western blotting). (b) Effect of NS50 on the expression of TNF-α-induced cell adhesion molecules (PCR). ✶ denotes P < 0.05 compared with cells treated with the blank control, # denotes P < 0.05 compared with cells treated with TNF-α, & denotes P < 0.05 compared with cells treated with TNF-α and SN50. (c) Effects of 1α,25-(OH)2D3 on NF-κB expression in HUVECs. (d) Effects of 1α,25-(OH)2D3 on NF-κB expression in HUVECs at different intervention times. ✶ denotes P < 0.05 compared with cells treated with the blank, # denotes P < 0.05 compared with cells induced with TNF-α, & denotes P < 0.05 compared with cells treated with 1α,25-(OH)2D3. TNF-α: Tumor necrosis factor-α, VDR-siRNA: Vitamin D receptor siRNA, 1α, 25(OH)2D3: 1α,25-dihydroxyvitamin D3, VCAM-1: Vascular cell adhesion molecule-1, PCR: Polymerase chain reaction, NF-κB: Nuclear factor kappa-B.

Article Snippet: [ ] To block NF-κB signaling, HUVECs were pretreated with the specific NF-κB inhibitor NF-κB SN50 (213546-53-3, MCE, USA) for 1 h and then incubated with 40 ng/mL TNF-α for 24 h. [ ] To assess NF-κB early activation, HUVECs were pretreated with 1α,25-(OH)2D3 for 30 min, followed by incubation with TNF-α for various periods as indicated.

Techniques: Expressing, Western Blot, Control, Polymerase Chain Reaction