Journal: The Journal of Biological Chemistry

Article Title: NF-κB Signaling Activation Induced by Chloroquine Requires Autophagosome, p62 Protein, and c-Jun N-terminal Kinase (JNK) Signaling and Promotes Tumor Cell Resistance *

doi: 10.1074/jbc.M116.756536

Figure Lengend Snippet: p62 is required for CQ-induced NF-κB activation. A and B, immunoblot analysis of p62 and GAPDH in Mel624 (A) and A431 (B) cells treated with or without CQ (25 μm) for 24 h. C and D, immunoblot analysis of p62 (C), p-IKK (D), IKK (D), and GAPDH in Mel624 cells transfected with control siRNA or siRNA targeting p62 (si-p62), followed by treatment with or without CQ (25 μm) for 8 h. E, luciferase reporter assay of NF-κB activity in WT and p62 knock-out (KO) MEF cells transfected with the reporter vector with specific NF-κB response elements (NF-κB-RE) followed by treatment with or without CQ (10 μm) for 18 h. F, luciferase reporter assay of NF-κB activity Mel624 cells transfected with si-control or si-p62 followed by transfection with the NF-κB-RE reporter vector and then treatment with or without CQ (25 μm) for 12 h. G, immunoblot analysis of p62 and GAPDH in A375 cells stably infected with sh-negative control (sh-NC) or sh-p62. H, immunoblot analysis of HIF-1α and GAPDH in sh-NC and sh-p62 A375 cells treated with or without CQ (25 μm) for 24 h. I, immunoblot analysis of p62 and β-actin in WT and p62 KO MEF cells treated with or without CQ (25 μm) for 24 h. J, immunoblot analysis of HIF-1α and β-actin in WT and p62 KO cells treated with the indicated concentrations of CQ for 2 h. The results were obtained from three independent experiments (mean ± S.D. (error bars), n = 3; *, p < 0.05 between comparison groups (Student's t test)).

Article Snippet: Luciferase Reporter Assays Cells were transfected with 1 μg of pGL3 AP-1-Luc , pGL3 CREB-Luc , or pGL3 NF -κ B-Luc and 0.025 μg of pRL- TK , which is used as a transfection efficiency control (Promega, Madison, WI), using GenJet TM Plus DNA in vitro transfection reagent (Signagen, Ijamsville, MD) according to the manufacturer's instructions.

Techniques: Activation Assay, Western Blot, Transfection, Luciferase, Reporter Assay, Activity Assay, Knock-Out, Plasmid Preparation, Stable Transfection, Infection, Negative Control

Journal: Cell Reports Medicine

Article Title: IL32 downregulation lowers triglycerides and type I collagen in di-lineage human primary liver organoids

doi: 10.1016/j.xcrm.2023.101352

Figure Lengend Snippet:

Article Snippet: RNAeasy Plus Mini kit , Qiagen , 74104.

Techniques: Recombinant, Protein Extraction, Staining, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Transfection, Gene Expression, Fluorescence, Software, Cell Counting