Journal: Molecules (Basel, Switzerland)

Article Title: Autonomous-Strengthening Adhesive Provides Hydrolysis-Resistance and Enhanced Mechanical Properties in Wet Conditions.

doi: 10.3390/molecules27175505

Figure Lengend Snippet: Scheme 1. Schematic of the proposed hydrophilic polymethacrylate-based adhesives with a rela- tively low crosslink density. Schematic illustrates the cascade of events resuming in the C1 and E1 copolymers that includes formation of copolymer network by free-radical polymerization and release of HEMA leachate when submerged in water. Note that the sol-gel-driven self-strengthening only resumes in water for E1 copolymer due to hydrolysis-polycondensation reaction at the trialkoxysilyl functional groups of MPS.

Article Snippet: Extracts of 0.4 mL were removed at 2, 3, and 4 days to analyze HEMA concentrations by HPLC (Shimadzu LC-2010C HT, software Labsolutions 5.93).

Techniques: Functional Assay

Journal: Cell Death & Disease

Article Title: Targeting KPNB1 overcomes TRAIL resistance by regulating DR5, Mcl-1 and FLIP in glioblastoma cells

doi: 10.1038/s41419-019-1383-x

Figure Lengend Snippet: a U87 and U251 cells were pretreated with CQ (40 μM) or Baf-A1 (5 nM) along with IPZ (16 μM) or IVM (16 μM) for 24 h and further with TRAIL (U87, 50 ng/ml; U251, 100 ng/ml) for 3 h. P62 in cell lysates was immunoprecipitated followed by western blot. b Representative images showed colocalization of p62, caspase-8 p43/p41/p18, and LC3 in U251 cells when treated with Baf-A1 (5 nM) and IPZ (16 μM) for 24 h and further with TRAIL (100 ng/ml) and z-DEVD-FMK (50 μM) for 8 h. Magnification, × 60; scale bar 25 μm. c U87 and U251 cells expressing shLC3B or not were treated with IPZ (16 μM) and CQ (40 μM) or Baf-A1 (5 nM) for 24 h and further with TRAIL (U87, 50 ng/ml; U251, 100 ng/ml) for 2 h. Then, cells were subjected to CHX pulse-chase assay by treating with CHX (U87, 20 μg/ml; U251 100 μg/ml) and z-VAD-FMK (20 μM) for indicated period of time. Representative images of western blot were shown in the upper panel. Quantification of grayscale ratio of cleaved caspase-8/α-tubulin by Photoshop software were shown in the lower panel. Results represent mean ± SD from two independent experiments. d , e U251 cells were pretreated as in a and further with z-IETD-FMK (20 μM) and TRAIL (100 ng/ml) for 6 h (western blot) or 24 h (flow cytometry), then subjected to flow cytometry ( d ) and western blot (e ). Results represent mean ± SD from three independent experiments. GAPDH and α-tubulin was used as loading controls. * P < 0.05

Article Snippet: Reagents and kits used in this study were listed below: importazole (IPZ) (Merck Millipore, Darmstadt, Germany), PMSF, propidium iodide (Sigma-Aldrich, St. Louis, USA), CalPhos Mammalian Transfection Kit (TaKaRa Bio, Kusatsu, Japan), protease inhibitor cocktail, RNase A, Click-iTTM Plus OPP Protein Synthesis Assay Kit Alexa FluorTM 488 picolyl azide (Thermo Scientific, Waltham, USA), phosphatase inhibitor, thiazolyl blue tetrazolium bromide (Sangon, Shanghai, China), cell lysis buffer for Western and IP, cycloheximide (CHX), nuclear and cytoplasmic protein extraction kit, RIPA (Beyotime, Nantong, China), LightCycler 480 SYBR Green I Master, protein A agarose, protein G agarose (Roche Diagnositics, Indianapolis, USA), bafilomycin A1 (Baf-A1), ivermectin (IVM), MG132, z-VAD-FMK (Selleck, Shanghai, China), Act D, chloroquine diphosphate (CQ), z-DEVD-FMK, z-IETD-FMK (Medchem Express, Monmouth Junction, USA), human recombinant TRAIL (PeproTech, Rocky Hill, USA), iScript Reverse Transcription Supermix (Bio-Rad, Berkeley, USA), immobilized 2′/3′-EDA-m 7 GTP (Jena Bioscience, Jena,Germany) were used in this study.

Techniques: Immunoprecipitation, Western Blot, Expressing, Pulse Chase, Software, Flow Cytometry