Journal: Cells

Article Title: Fabrication Scaffold with High Dimensional Control for Spheroids with Undifferentiated iPS Cell Properties

doi: 10.3390/cells12020278

Figure Lengend Snippet: Fluorescent immunostaining results of the multidifferentiation abilities. Blue coloration shows where the cells localize, and they emit green or red (FOXA2, SOX17, PAX2, cardiac Troponin T, OTX2, OCT4, β tubulin) depending on the localization of each protein.

Article Snippet: MA, USA), mouse monoclonal anti-TRA-1-81 (1:1000; 4745, CST), mouse monoclonal anti-SSEA4 (1:1000; 4755, CST), rabbit anti-NANOG (1:500; 4903, CST), rabbit monoclonal anti-FOXA2/HNF3β (1:400; 8186S, CST), and mouse monoclonal anti-SOX17 (1:100; TA500044, Origene, Rockville, MD, USA) as early endoderm markers, rabbit polyclonal anti-Cardiac Troponin T (1:100; 15513-1-AP, Proteintech, Rosemont, IL, USA) and mouse monoclonal anti-PAX2 (1:100; MBS4380650, MyBioSourse, San Diego, CA, USA) as early ectoderm markers, and goat polyclonal anti-OTX2 (1:100; SC031B, R & D systems) and rabbit monoclonal anti-β Tubulin (1:100; 80713-1-RR, Proteintech) as early ectoderm markers.

Techniques: Immunostaining

Journal: Cell Death & Disease

Article Title: NIT1 suppresses tumour proliferation by activating the TGFβ1–Smad2/3 signalling pathway in colorectal cancer

doi: 10.1038/s41419-018-0333-3

Figure Lengend Snippet: a Overexpression or knockdown of NIT1 in CACO2 and SW620 cells analysed by western blotting. β-tubulin was used as a loading control. b Effects of NIT1 on cell proliferation were determined by CCK-8 cell proliferation assays. The data are represented as the means ± SD of five samples. c Effects of NIT1 on cell proliferation were determined by colony formation assays. The data are represented as the means ± SD of three samples

Article Snippet: The membrane was then blocked in PBST solution containing 5% non-fat milk and was incubated at 4 °C overnight with the specific primary antibodies anti-NIT1 (1:500 dilution; Proteintech, USA), anti-β-tubulin (1:2000 dilution; zsgb-bio, Beijing), anti-GAPDH (1:5000 dilution; Proteintech, USA), and anti-p53 (1:500 dilution; Proteintech, USA), anti-p21 (1:1000 dilution; Abcam, USA), anti-p27 (1:1000 dilution; Abcam, USA), anti-cyclinD1 (1:1000 dilution; Abcam, USA), anti-CDK4 (1:500 dilution; Proteintech, USA), anti-CDK6 (1:500 dilution; Proteintech, USA), anti-myc (1:500 dilution; Proteintech, USA), anti-bax (1:1000 dilution; Abcam, USA), anti-bcl2 (1:1000 dilution; Abcam, USA), anti-caspase3 (1:1000 dilution; CST, USA), anti-PARP (1:1000 dilution; CST, USA), anti-SARA (1:500 dilution; Abcam, USA), anti-Smad2/3 (1:500 dilution; Abcam, USA), anti-Smad2 (1:1000 dilution; Abcam, USA), anti-Smad3 (1:1000 dilution; Abcam, USA), anti-p-Smad2 (1:500 dilution; CST, USA), and anti-p-Smad3 (1:1000 dilution; CST, USA), followed by incubation with their respective appropriate second antibodies.

Techniques: Over Expression, Western Blot, CCK-8 Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: New melanocortin-like peptide of E. coli can suppress inflammation via the mammalian melanocortin-1 receptor (MC1R): possible endocrine-like function for microbes of the gut

doi: 10.1038/s41522-017-0039-9

Figure Lengend Snippet: Antibody to MC1R blocks suppressant effect of MECO-1 and alpha-MSH. Murine macrophage-like RAW 264.7 cells were pretreated with specific antibodies (“anti-MC1R Ab”) against melanocortin-1 receptor (MC1R) for 10 min prior to addition of HMGB1 in the absence or presence of MECO-1 or alpha-MSH at 10 −12 M. At 6 h, the TNF content of the cell-free medium was determined by ELISA, and expressed as mean ± SEM of two independent experiments performed in duplicate. ACTH at the same concentration gave results that were indistinguishable from those of alpha-MSH and MECO-1 (data not shown)

Article Snippet: Final LPS content was determined using the Limulus amebocyte lysate assay (BioWhittaker Inc.) as described previously., LPS ( E. coli , 0111:B4) and macrophage colony-stimulating factor were purchased from Sigma Chemical Co. Anti-MC1R antibody was purchased from (MC1R Antibody (N-19): sc-6875) Santa Cruz Biotechnology Inc.

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: New melanocortin-like peptide of E. coli can suppress inflammation via the mammalian melanocortin-1 receptor (MC1R): possible endocrine-like function for microbes of the gut

doi: 10.1038/s41522-017-0039-9

Figure Lengend Snippet: Agouti (endogenous MC1R inverse agonist) inhibits the effects of MECO-1 and alpha-MSH on endotoxin-stimulated TNF release

Article Snippet: Final LPS content was determined using the Limulus amebocyte lysate assay (BioWhittaker Inc.) as described previously., LPS ( E. coli , 0111:B4) and macrophage colony-stimulating factor were purchased from Sigma Chemical Co. Anti-MC1R antibody was purchased from (MC1R Antibody (N-19): sc-6875) Santa Cruz Biotechnology Inc.

Techniques: Control

Journal: NPJ Biofilms and Microbiomes

Article Title: New melanocortin-like peptide of E. coli can suppress inflammation via the mammalian melanocortin-1 receptor (MC1R): possible endocrine-like function for microbes of the gut

doi: 10.1038/s41522-017-0039-9

Figure Lengend Snippet: a Structure of melanocortins and similar peptides. The four common melanocortins of mammals are derived from one pro-opiomelanocortin (POMC) precursor. ACTH and alpha-MSH are products of alternative processing of the same precursor and have very similar interactions with four of the melanocortin receptors. (The exception is MC2R; in the adrenal MC2R, is the dominant receptor and enables ACTH to have an effect that is orders of magnitude greater than that of alpha-MSH.) While the primary sequences of amino acids 1–13 are identical in ACTH and alpha-MSH, the latter has two post-translational moieties, an N-terminal acetyl and a C-terminal amide, that are important for full potency of alpha-MSH. That MECO-1 lacks the HFRW canonical sequence of the mammalian melanocortins but has bioactivity at MC1R equal to alpha-MSH was a surprise to us. Do note that three of the four substitutions are considered “conservative”. b Mitochondrial peptides. The C-termini of the two elongation factors of human mitochondria are compared to MECO-1. Note that each of them is about as close to MECO-1 as each is to the other. c Microbial peptides. The first eight sequences are those with the closest matches with MECO-1. The last two are from Bacteriodetes, organisms that are typically highly represented in the human colon

Article Snippet: Final LPS content was determined using the Limulus amebocyte lysate assay (BioWhittaker Inc.) as described previously., LPS ( E. coli , 0111:B4) and macrophage colony-stimulating factor were purchased from Sigma Chemical Co. Anti-MC1R antibody was purchased from (MC1R Antibody (N-19): sc-6875) Santa Cruz Biotechnology Inc.

Techniques: Derivative Assay, Sequencing