Journal: Frontiers in Physiology

Article Title: Yes-Associated Protein 1 Plays Major Roles in Pancreatic Stellate Cell Activation and Fibroinflammatory Responses

doi: 10.3389/fphys.2019.01467

Figure Lengend Snippet: YAP siRNA decreases PaSC expansion in culture and alters expression of genes involved in matrix remodeling. imPaSC were transfected with nontargeting negative control ( NT , 10 nmol/L) or a target siRNA ( YAP , 10 nmol/L) mixed with lipofectamine RNAiMAX, and 24 h later cells were used for experiments. (A) Graph shows quantitation of YAP mRNA expression relative to negative control at 48 h post-transfection. Data are mean ± SD, n = 3; * p < 0.05 vs. NT. (B) MTT assay for YAP siRNA-treated cells relative to NT. (C) The graph shows imPaSC numbers counted 0 and 48 h after treatment with either NT or YAP siRNA. The cell numbers were assessed by trypan blue exclusion using a hemocytometer, and expressed as a fold increase over time 0 (1.5 × 10 5 ). (D) PI uptake was measured in cells treated with siRNA to detect any disrupted plasma membrane integrity. After incubation, a portion of cells was permeabilized for 5 min with 0.1% Triton X100 to establish a positive control. Data in graphs (B–D) are mean ± SD, n = 3–6 independent experiments; * p < 0.05 vs. NT. shows protein levels of caspase-3 as a marker of caspase-dependent apoptosis . (E) Graph shows mRNA levels of the indicated genes in YAP siRNA treated cells compared to NT. Data is mean ± SD, n = 5.

Article Snippet: Silencer nontargeting negative control (10 nmol/L; Mock transfection) or YAP siRNA (10 nmol/L) were mixed with Lipofectamine RNAiMAX (#13778075 ThermoFisher Scientific) according to the manufacturer’s recommendations and added to the cells.

Techniques: Expressing, Transfection, Negative Control, Quantitation Assay, MTT Assay, Clinical Proteomics, Membrane, Incubation, Positive Control, Marker

Journal: Biochemistry and Biophysics Reports

Article Title: Characterization of an African trypanosome mutant refractory to lectin-induced death

doi: 10.1016/j.bbrep.2015.08.013

Figure Lengend Snippet: (A) N-glycan and GPI biosynthetic precursors analysis of parental and 3B2 cells treated with or without tunicamycin (Tun), a Dol-PP-GlcNAc biosynthesis inhibitor. GDP-[ 3 H]Man-labeled products from a cell-free biosynthesis system were fractionated by TLC, and detected by autoradiography. Dol-P-Man, dolichol phosphoryl Man; PP1, ethanolamine phosphate Man 3 GlcN-acylinositol lyso- phosphatidylinositol; and PP3, ethanolamine phosphate Man 3 GlcN-acylinositol alkyl-acyl-phosphatidylinositol, GPI precursors; O, origin. (B) Fla1p western blot. Membrane proteins from parental and clone 3B2 were either mock treated or incubated with 1U Endo H and then probed with anti-Fla1p serum.

Article Snippet: Fla1p was detected from 2×10 7 cell equivalents that were either mock treated or incubated with 1U Endo H (24 h, 37 °C, 500 U/mL, New England Biolabs, Beverly MA).

Techniques: Labeling, Autoradiography, Western Blot, Incubation