Journal: The Journal of Biological Chemistry

Article Title: Small molecule LATS kinase inhibitors block the Hippo signaling pathway and promote cell growth under 3D culture conditions

doi: 10.1016/j.jbc.2022.101779

Figure Lengend Snippet: GA-017 promotes growth and/or organoid formation of SKOV3 and A431 spheroids, MDCK cysts, and mouse intestinal organoids. A and B , SKOV3 cells were cultured in wells of low-attachment U-bottom plates with DMSO or GA-017 for up to 6 days. A , images of SKOV3 cells cultured for 4 days in the well. Left : control (DMSO), right : 10 μM GA-017. The bars represent 500 μm. B , time course of SKOV3 cell growth evaluated using an ATP assay. Each RLU was normalized by that on day 0. DMSO: vehicle control, GA-017: 10 μM GA-017. The data represent means ± SD of three independent experiments. ∗ p <0.01. C and D , A431 cells were cultured in hanging drops with DMSO or 10 μM of GA-017 for 2 days. C , images of A431 cells cultured in the drop. Left : control (DMSO), right : 10 μM GA-017. D , the number of cells cultured with DMSO or GA-017. Y-axis indicates relative cell number (% of control) to vehicle control (DMSO). The data represent means ± SD of 11 independent experiments. ∗ p <0.01. E and G , MDCK cells were cultured in Matrigel for 6 days with DMSO (0.01%) or 10 μM GA-017. E , fluorescent images of MDCK cysts cultured for 6 days. Upper : control (DMSO), lower: 10 μM GA-017. β-catenin: magenta , phalloidin: green , nuclear (DAPI): blue . F , total number of MDCK cysts (diameter >30 μm). The data represent means ± SD of three independent experiments. ∗ p <0.05. G , size distribution of MDCK cysts formed in Matrigel with GA-017 or DMSO. Shown is the ratio to the total cyst number. H–K , mouse intestinal organoids were cultured in Matrigel for up to 9 days with DMSO (0.01%) or 10 μM GA-017. H , images of organoids cultured for 9 days in Matrigel. Left : control (DMSO), right : 10 μM GA-017. I , time course of organoid formation. Y-axis indicates the total volume of organoids/total volume of initial crypts (day 1). ∗ p <0.05, ∗∗ p <0.01. J , size distribution of intestinal organoids formed in Matrigel with GA-017 or DMSO. Shown is the ratio to the total organoid number. K , relative Lgr5 , Vil1 , and Alpi mRNA expression in organoids on day 9, normalized by Gapdh . The data represent means ± SD of three independent experiments. Statistical significance was analyzed using Dunnett’s test for B or Student’s t test for D, F, I, and K. ∗ p <0.01. DMSO, dimethyl sulfoxide; MDCK, Madin-Darby canine kidney; NS, not significant; NT, nontreatment.

Article Snippet: Cold Matrigel Matrix GFR (Corning Incorporated) was spread on a 24-well plate at 50 μl per well and allowed to sit at 37 °C for 15 min. MDCK cell suspension (1.1 × 10 4 cells/ml) in the indicated medium including 0.2% Matrigel Matrix GFR was dispensed to the wells at 900 μl/well.

Techniques: Cell Culture, Control, ATP Assay, Expressing

Journal: Cell

Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

doi: 10.1016/j.cell.2018.08.067

Figure Lengend Snippet:

Article Snippet: Matrigel Basement Membrane Matrix Growth Factor Reduced (GFR) Phenol Red-Free LDEV-Free , Corning , Cat#356231.

Techniques: Recombinant, Coagulation, Antibody Labeling, Membrane, Plasmid Preparation, Polymer, Blocking Assay, Staining, Imaging, cDNA Synthesis, RNAscope, DNA Library Preparation, Software, Hybridization

Journal: International Journal of Molecular Sciences

Article Title: Engineered Mesenchymal Stem Cells Expressing Stromal Cell-derived Factor-1 Improve Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats

doi: 10.3390/ijms19123730

Figure Lengend Snippet: ( A ) Microscopic fields showing migrated HUVECs at the bottom of the PET membrane; ( B ) Bar graph representing the percentage of migrated HUVECs per migrated field in response to BM-MSCs and SDF-1 eMSCs. SDF-1 eMSCs had a greater chemotactic effect than BM-MSCs. Original magnification: ×100; ( C ) Fluorescence microscopy images showing the formation of the tube network on the Matrigel matrix. BM-MSC- and SDF-1 eMSC-treated groups had greater capillary-like structures than EBM-treated groups; ( D ) Quantitative results for total meshes, total nodes, and total junctions in each group. Original magnification: ×50. * p < 0.05 compared with the DM ED group. ** p < 0.01 compared with the DM ED group.

Article Snippet: A total of 200 μL of BD Growth Factor Reduced (GFR) Matrigel Basement Membrane Matrix (BD Biosciences) was added to a 24-well plate and incubated at 37 °C for 30 min. HUVECs (1 × 10 5 cells/well) were incubated on a plate coated with Matrigel in endothelial growth media (EGM), endothelial basal media (EBM) (no growth supplement), and EBM with 30% conditioned media (BM-MSC and SDF-1 eMSC).

Techniques: Fluorescence, Microscopy

Journal: Cell reports

Article Title: GPCR kinases generate an APH1A phosphorylation barcode to regulate amyloid-β generation

doi: 10.1016/j.celrep.2022.111110

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Matrigel GFR matrix , Fisher Scientific , Cat# 354230.

Techniques: Recombinant, PathHunter β-Arrestin Assay, Mutagenesis, BIA-KA, Software, Membrane