Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation

doi: 10.1186/s13046-019-1317-6

Figure Lengend Snippet: DU145 cells adhesion, invasion and intracellular activated pathways upon exposure to LO145 or LOR80. a DU145 adhesion, expressed as % of adherent cells, upon exposure to LO145 or LOR80 vs vehicle (considered as 100% control) and evaluated both in absence or presence of vitronectin (VN) for either 30 or 60 min. Stat. significances are expressed on vehicle. b DU145 adhesion measured as above, on VN after 60′ of exposure to LO145 or LOR80, untreated or pre-treated for 45′ RT with anti-αV-integrin monoclonal blocking antibody (anti-αV Ab) and indicated as LO145 αv and LOR80 αv , respectively; as negative control cells were treated by the anti-αV Ab alone. (LO145 αv / LO145)**; (LOR80 αv / LOR80)***; (LOR80/ vehicle) ***. c Western blotting analysis of pFAK, FAK, pAKT, AKT and γ-tubulin on cell lysates from DU145 untreated (CTR), or treated with LO145 or LOR80, alone or pre-exposed to anti-αV Ab (see above). pFAK and pAKT quantification is also reported. d Adhesion of DU145 measured as above, untreated (considered as 100%) or pretreated with the AKT inhibitor GDC-0068 for 16 h, were exposed to vehicle, LO145 or LOR80. (GDC-0068/ vehicle)***; (LOR80/ vehicle) **; (GDC-0068+ LOR80/ LOR80)* ( e ) DU145 invasion, expressed as % of invading cells, upon 16 h exposure to LOR80 treated with anti-αV Ab (LOR80 av ) or control Ab (LOR80 CTR Ab ) vs vehicle (considered as 100% control); as negative control cells were treated by CTR Ab alone (anti-pAKT antibody ineffective on non-permeabilized intact cells); (LOR80/ vehicle)***; (LOR80 CTR Ab / vehicle)***; (LOR80 av / LOR80)** ( f ) Invasion of DU145 measured as above, untreated (considered as 100%), or pretreated with the AKT inhibitor GDC-0068 for 6 h and exposed to vehicle, LO145 or LOR80;(GDC-0068/ vehicle)***; (LOR80/ vehicle)***; (GDC-0068 + LOR80/ LOR80)***. Results are representative of a single experiment and several experiments yielded similar results ( b ), or are the average of at least two experiments ( a ; d ; e ; f )

Article Snippet: Cell adhesion assays were performed using 24-well tissue culture plates uncoated/coated with vitronectin (VN) (Corning, NY 14831 USA).

Techniques: Control, Blocking Assay, Negative Control, Western Blot

Journal: Biomacromolecules

Article Title: Squaramide-Based Supramolecular Materials for Three-Dimensional Cell Culture of Human Induced Pluripotent Stem Cells and Their Derivatives

doi: 10.1021/acs.biomac.7b01614

Figure Lengend Snippet: 3D cell culture of hiPSCs in hydrogel 3 (3.1 mM): Representative images at ∼24 (A) and ∼72 h (B). Dead cells were detected using the NucGreen Dead reagent. Scale bar: 100 μm. (C) Distribution of spheroid diameters in a hydrogel of 3 (3.1 mM) after 24, 48, and 72 h; ∼135 spheroids per day were measured. (D) FACS analysis of single cells of hiPSCs cultured under standard conditions on a vitronectin surface in 2D ( left control, middle with TRA-1-60 and SSEA-4 antibodies) or after retrieval from hydrogel 3 (3.1 mM) after 24 h of culture ( right with TRA-1-60 and SSEA-4 antibodies). Upper panels : side scatter (SSC-A) and forward scatter (FSC-A) showing live cell population (black circles are the gated live cells); expression levels of TRA-1-60 ( middle panels ) and SSEA-4 ( lower panels ) pluripotent stem cell markers of the gated live cells.

Article Snippet: A hiPSC line (SFLB6) was maintained on human recombinant vitronectin-coated plates in TeSR-E8 cell culture medium (STEMCELL Technologies). hiPSC-derived endothelial cells (ECs) were cultured on gelatin-coated plates in EC-SFM (Invitrogen) additionally supplemented with 1% platelet-poor serum, 30 ng/mL of VEGF, and 20 ng/mL of bFGF as previously described.

Techniques: Cell Culture, Expressing