Journal: Cell

Article Title: Inadequate DNA damage repair promotes mammary transdifferentiation leading to BRCA1 breast cancer

doi: 10.1016/j.cell.2019.06.002

Figure Lengend Snippet: KEY RESOURCES TABLE (antibodies and reagents)

Article Snippet: Sorted YFP + MECs were embedded in 15-20 μl Matrigel (Corning, 356231) in a 48-well plate, allowed to solidify for 5-10 mins, and then overlaid with DMEM/F12 advance media, supplemented with HEPES (1:100, GibcoTM, 15630080), GlutaMAX (1:100, GibcoTM, 35050-061), B27 (1:50, Thermo Fisher, 17504044), 100 ng/ml A83-01 (Tocris Bioscience, 2939), 50 ng/ml EGF (PeproTech, AF-100-15), 100 ng/ml Noggin (PeproTech, 120-10C), 500 ng/ml commercial R-spondin1 (Peprotech, 120-38) or conditioned R-spondin1 medium from RSPO cells (Cultrex, 3700-100-01), 100 ng/ml Nrg1 (R&D, 396-HB), 10 μM Y-27632 (ROCK inhibitor, Sigma Aldrich, Y0503) which is only necessary when organoids are first prepared, thawed, or when organoids are dissociated to single cells.

Techniques: Recombinant, Mutagenesis, Expressing, Software, Multiplex Assay

Journal: eLife

Article Title: A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction

doi: 10.7554/eLife.44530

Figure Lengend Snippet: ( A ) Western blot images of neuregulin 1-β1 (NRG1- β1; top) and β-actin (bottom) protein expression in three-independent ESC-derived MN cultures (N1 – N3). ( B ) Bar graph quantifying qRT-PCR results for epsilon subunit of AChR (CHRNE) gene expression in 2D (light blue) and 3D (blue bars) muscle alone, NRG1- β1 treated muscle cultures, and neuromuscular co-cultures. N = 3 muscle patient donors for data presented in ( B ). In ( B ) each symbol represents data from one muscle patient donor. Values in ( B ) are mean ±SEM. *p<0.05 and **p<0.01 compared to 3D muscle alone, and ## p<0.01 compared to 2D muscle culture. ( C ) Representative GCaMP6 epifluorescence images of 3D neuromuscular co-cultures pre-treated for 3 days with healthy (top) or myasthenia gravis (MG, bottom) patient IgG, together with human complement, and then stimulated with ACh (100 μM). Scale bars, 250 μm. ( D ) Bar graph indicating the percent total 3D tissue area occupied by muscle fibers responding to acetylcholine (ACh) stimulation in healthy and MG patient IgG treated neuromuscular co-cultures. n = 4 neuromuscular tissues treated with healthy IgG and three neuromuscular tissues each treated with serum IgG from one of three separate MG patient donors. Values in ( D ) are mean ±SEM. ***p<0.001.

Article Snippet: In experiments using neuregulin1-β1 treatment, recombinant human neuregulin1-β1 (R and D Systems) was supplemented in the culture media at 5 nM.

Techniques: Western Blot, Expressing, Derivative Assay, Quantitative RT-PCR

Journal: eLife

Article Title: A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction

doi: 10.7554/eLife.44530

Figure Lengend Snippet: List of primary antibodies.

Article Snippet: In experiments using neuregulin1-β1 treatment, recombinant human neuregulin1-β1 (R and D Systems) was supplemented in the culture media at 5 nM.

Techniques: