Journal: Cell Biology and Toxicology
Article Title: The COP9 signalosome stabilized MALT1 promotes Non-Small Cell Lung Cancer progression through activation of NF-κB pathway
doi: 10.1007/s10565-024-09888-z
Figure Lengend Snippet: CSN5 interacted with MALT1 to activate NF-κB signaling pathway. A Gene Ontology categories by Gene Ontology analysis of genes interacted with MALT1 in A549 cells. BP biological process, MF molecular function, CC cellular component. The COP9 signalosome was highlighted. B In A549 cells, CSN5 protein in MALT1 Co-IP protein mix detected by MS, arrow indicated the identified CSN5 peptide peak. C The interaction between CSN5 and MALT1 in A549 cells was detected by Co-IP assays. D-F The effects of MALT1 ( D ), MI-2 ( E ) and CSN5 ( F ) on NF-κB signaling pathway activation in A549 cells were detected by immunoblotting. G-H Dual-luciferase reporter assays were used to analyze NF-κB activation in A549 cells after transfection ( G, I ) or MI-2 treatment ( H ). J Rescue assays were performed with dual-luciferase reporter assays to detected the NF-κB activation in A549 cells. Each experiment was performed in triplicate and data are presented as mean ± SD. One-way ANOVA, Dunnett’s Multiple comparison test and LSD multiple comparison test were used to analyze the data (* p < 0.05, ** p < 0.01, *** p < 0.001)
Article Snippet: Protein was extracted in cell lysis buffer (Beyotime, Shanghai, China) containing protease inhibitor (SelleckChem), separated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and blotted onto PVDF membranes (Millipore, MA, USA), which incubated with primary antibodies against MALT1 (CST, 2494, Danvers, USA), CSN5 (Proteintech, 27511–1-AP), FBXO3 (Proteintech, 17803–1-AP), p-p65 (Absci, AB11014, OR, USA), Ub (Proteintech, 10201–2-AP), β-actin (Santa Cruz, 47778, CA, USA), and GAPDH (Santa Cruz, 47724).
Techniques: Co-Immunoprecipitation Assay, Activation Assay, Western Blot, Luciferase, Transfection, Comparison