gsk2850163  (MedChemExpress)

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Brucella BvrR activates ATF2/IL‐6 and NF‐κB p65/TNF‐α signaling via IRE1. (A) Transmission electron microscopy analysis of ER morphology in HMC3 cells. (B) Western blot analysis of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α protein levels. (C–G) Quantification of p‐IRE1, p‐ATF2, p‐p65, IL‐6, and TNF‐α expression normalized to GAPDH. Data are presented as means ± standard deviations from at least three independent experiments; * p < 0.05 versus Control; # p < 0.05 versus IXA4; & p < 0.05 versus GSK2850163; ! p < 0.05 versus BvrR.

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Control

Journal: MicrobiologyOpen

Article Title: BvrR From Brucella abortus Induces Neuroinflammation Through IRE1‐Mediated Activation of ATF2 and NF‐κB

doi: 10.1002/mbo3.70219

Figure Lengend Snippet: Effect of BvrR, IXA4, and GSK2850163 on IL‐6 and TNF expression. (A) ELISA measurement of IL‐6 in supernatants. (B) ELISA measurement of TNF‐α in supernatants. (C) RT‐qPCR analysis of IL6 mRNA expression. (D) RT‐qPCR analysis of TNF mRNA expression. Each assay was performed in six replicates, and data are presented as means ± standard deviations; * p < 0.05, ** p < 0.01 versus Control; # p < 0.05, ## p < 0.01 versus IXA4; & p < 0.05, && p < 0.01 versus GSK2850163; ! p < 0.05, !! p < 0.01 versus BvrR.

Article Snippet: The IRE1 activator IXA4 (#145279) and inhibitor GSK2850163 (#40776) were acquired from MedChemExpress (New Jersey, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Biomolecules

Article Title: Cardiopulmonary Bypass-Induced IL-17A Aggravates Caspase-12-Dependent Neuronal Apoptosis Through the Act1-IRE1-JNK1 Pathway

doi: 10.3390/biom15081134

Figure Lengend Snippet: IL-17A enhances OGD/R-induced apoptosis through the Act1-IRE1-JNK1 pathway. ( a – e ) Apoptosis- and ERS-related protein expression levels in different B104 cell groups. ( a ) Typical Western blot bands showed that compared with the Concel group, the hydrolysis levels of Caspase-3 ( b ) and Caspase-12 ( c ) in B104 cells treated with OGD/R were significantly increased, and the phosphorylation levels of IRE1 ( d ) and JNK1 ( e ) were also significantly increased. On this basis, the hydrolysis levels of Caspase-3 and -12 and the phosphorylation levels of IRE1 and JNK1 were further increased after the intervention of rmIL-17A. After intervention with GSK2850163, the above rising trend was significantly suppressed. * p < 0.05 compared to the Concel group; # p < 0.05 compared to the OGD/R++DMSO+ Vehicel group; & p < 0.05 compared to the OGD/R+DMSA+rmIL-17A group, n = 6 per group; ( f , g ) TUNEL staining results of B104 cell in different groups: compared with the Normoxia group, the number of apoptotic cells in B104 cell treated with OGD/R was significantly increased, and rmIL-17A could significantly increase the apoptosis induced by OGD/R. After the intervention of GSK2850163, the increase in apoptosis induced by rmIL-17A and OGD/R was significantly reversed. * p < 0.05 compared to the Normoxia+Vehicel group; # p < 0.05 compared to the OGD/R+Vehicel group; & p < 0.05 compared to the OGD/R+rmIL-17A group, n = 6 per group. ( h ) At the cellular level, after OGD/R treatment, the binding of Hsp90 to Act1 increased, but the binding of HSP90 to IRE1 decreased. After intervention with GSK2850163, the binding of Hsp90 to Act1 decreased, while the binding of HSP90 to IRE1 increased.

Article Snippet: For OGD induction, cultures were transferred to glucose-free DMEM and placed in a tri-gas incubator pre-equilibrated to 2% O 2 , 5% CO 2 , and 93% N 2 for 1 h. Reperfusion was initiated by replacing the medium with complete growth medium (DMEM + 10% fetal bovine serum) under normoxic conditions (21% O 2 , 5% CO 2 , 74% N 2 ) for 24 h. Pharmacological interventions—recombinant IL-17A (250 ng/mL; RP-8621, Thermo Fisher Scientific, Waltham, MA, USA) and the IRE1α-specific inhibitor GSK2850163 (HY- U00459 , MedChemExpress, Monmouth Junction, NJ, USA)—were administered at OGD initiation.

Techniques: Expressing, Western Blot, Phospho-proteomics, TUNEL Assay, Staining, Binding Assay