Journal: Cell Death & Disease

Article Title: TNFAIP8 regulates autophagy, cell steatosis, and promotes hepatocellular carcinoma cell proliferation

doi: 10.1038/s41419-020-2369-4

Figure Lengend Snippet: a Fifty micrograms of lysates from HCC cells immunoblotted with indicated antibodies. b Fifty micrograms liver cancer tissue lysates with different stages of liver cancer were immunoblotted with indicated antibodies. c HepG2, SK-Hep1 cells were transfected with EV or TNFAIP8-Myc plasmid for 30 h, and cell lysates from transient and stable HepG2-TNFAIP8 expressing cells were immunoblotted with indicated antibodies. d HepG2 cells were grown in complete medium or serum-free (S.F.) medium for 24 h and then transfected with EV or TNFAIP8-Myc plasmid for 30 h. Fifty micrograms of proteins were immunoblotted. e HepG2 cells were transfected with control siRNA, or TNFAIP8 siRNA (100 nM) for 30 h and lysates were immunoblotted with autophagy related bio-markers. f HepG2 cells were grown in complete medium or serum-free (S.F.) medium for 24 h and then transfected with control siRNA or TNFAIP8 siRNA (100 nM) for 30 h as indicated, and lysates were immunoblotted with indicated antibodies. g HepG2 cells were pretreated with the autophagy inhibitor, 3-MA for 18 h and transfected with EV and TNFAIP8-Myc plasmid for 24 h in the presence of 3-MA and cell lysates were immunoblotted with indicated antibodies. h , i Effect of TNFAIP8 overexpression and 3-MA on LC3β related puncta formation in HepG2 cells were analyzed by Cyto-ID green fluorescence staining ( h ), and number of puncta were measured from TNFAIP8-transfected or 3-MA pretreated and TNFAIP8-transfected cells and plotted ( i ). *** P < 0.001 relative to TNFAIP8 transfected. EV: empty vector, Comp. medium: complete medium, S.F. medium: serum-free medium, N: normal, T: tumor. Arrow indicates ATG3 specific band.

Article Snippet: TNFAIP8-Myc-DDK-tagged ORF cDNA plasmid (Cat # RC202729) was obtained from Origene (Rockville, MD), TNFAIP8-his tagged protein (Cat # 14559-H07E) from Sino Biological (Wayne, PA), TNFAIP8 siRNA (Cat # L-020589), control siRNA (Cat # D-001830) from Dharmacon (Lafayette, CO), Oil Red O stain (Cat # O0625 from Sigma (St. Louis, MO), CYTO-ID® Autophagy Detection Kit (Cat # ENZ-51031-0050) from Enzo Life Science (Farmingdale, NY), G418 (Geneticin) (Cat # 11811-031) from Gibco (Gaithersburg, MD), MTT reagent (Cat # 102227) from MP Biomedical (Solon, OH), CellRox green reagents (Cat # C10444) from Molecular Probes, regorafenib (Cat # R8024) from LC Laboratories, sorafenib (Cat # S1040) from Selleckchem, HCC-TMA slide (Cat # BC03116) from US Biomax (Rockville, MD), TMA-Alcohol Steatosis (Cat # TMA.AS 1810055) and TMA No Alcohol Steatohepatitis (Cat # TMA.

Techniques: Transfection, Plasmid Preparation, Expressing, Over Expression, Fluorescence, Staining

Journal: Cell Reports Medicine

Article Title: Single-Cell Analyses Identify Dysfunctional CD16 + CD8 T Cells in Smokers

doi: 10.1016/j.xcrm.2020.100054

Figure Lengend Snippet:

Article Snippet: Cell-ID Intercalator-Ir—500 μM , Fluidigm , Cat# 201192B.

Techniques: Binding Assay, Recombinant, Modification, Saline, Staining, Multiplex Assay, Hybridization, DNA Methylation Assay, RNA Sequencing, Microarray, Software