Journal: Cell reports

Article Title: Activity-assembled nBAF complex mediates rapid immediate early gene transcription by regulating RNA polymerase II productive elongation

doi: 10.1016/j.celrep.2024.114877

Figure Lengend Snippet: (A) Schematic representation of three biochemically distinct BAF complexes: nBAF (neuronal cBAF), GBAF, and PBAF. Complex-defining unique subunits are represented in colors. Subunits shared by all three complexes are not indicated. Degraders and small-molecule inhibitors to target each complex are noted below. Schematic was created using BioRender. (B) Neurons were treated with DMSO (control) or the indicated concentrations of ACBI1 or cis -ACBI1 (inactive isomer of ACBI1) for 3 h. Whole-cell lysates were electrophoresed, western blotted, and probed for the indicated BAF subunits. (C–E) Neuronal lysates were used to immunoprecipitate SMARCA4 (C), ARID1A (D), and ARID1B (E). Five percent of the cell lysate was used as input. Histone 3 and IgG are depicted as loading controls. (F and G) Transcriptional assays in which Arc pre-mRNA normalized by GAPDH pre-mRNA levels is illustrated. (F) Neurons were treated with ACBI1 or cis -ACBI1 for 3 h followed by bicuculline and 4AP treatment for 15 min (Bic + 4AP). (G) Neurons were treated with BAF ATPase domain inhibitors BRM014 and FHT for 30 min, followed by Bic + 4AP treatment to induce neuronal activity. Gray dots represent biological replicates, error bars show SE of the mean. * p < 0.05 and ** p < 0.01. One-way ANOVA was followed by Tukey’s post hoc test. Approximate position of the nearest molecular weight marker is depicted against each band.

Article Snippet: To inhibit BAF ATPase function, BRM014 (compound 14, MedChemExpress; catalog no. HY-119374) and FHT-1015 (MedChemExpress; catalogue no. HY-144896) were used at concentrations described in figure legends.

Techniques: Control, Western Blot, Activity Assay, Molecular Weight, Marker

Journal: Cell reports

Article Title: Activity-assembled nBAF complex mediates rapid immediate early gene transcription by regulating RNA polymerase II productive elongation

doi: 10.1016/j.celrep.2024.114877

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: To inhibit BAF ATPase function, BRM014 (compound 14, MedChemExpress; catalog no. HY-119374) and FHT-1015 (MedChemExpress; catalogue no. HY-144896) were used at concentrations described in figure legends.

Techniques: Recombinant, Plasmid Preparation, Magnetic Beads, One Step RT-PCR, SYBR Green Assay, Mass Spectrometry, Software

Journal: International Journal of Bioprinting

Article Title: Blood-derived biomaterials for tissue graft biofabrication by solvent-based extrusion bioprinting

doi:

Figure Lengend Snippet: 3D bioprinting of specific tissue constructs using bioinks functionalized with blood-derived biomaterials

Article Snippet: Kumar et al. (2019); ACS Biomater Sci Eng [ ] , 1 mL of the bioink: 990 μL of fibrinogen solution (60 mg/mL), 155 mg of gelatin- furfuryl at 25°C Heat it at 37 °C for 1 h with stirring, add Rose Bengal (10 μL) Polymerization: 50 units/mL thrombin, and 80 mM CaCl 2 , Human-induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CM) Human CM AC 16 cell lines (ATCC) , Fibrinogen Thrombin (#) , SEM, swelling assay, rheology, Fourier transform infrared spectroscopy (FTIR), biocompatibility Cardiac cell-laden constructs To validate the applicability of this scaffold toward mimicking cardiac tissue.

Techniques: Construct, Formulation, Migration, In Vivo, Clinical Proteomics, In Vitro, Micro-CT, Concentration Assay, Gene Expression, Transplantation Assay, Activation Assay, Immunohistochemistry, Immunohistochemical staining, Fourier Transform Infrared Spectroscopy, Spectroscopy, Functional Assay, In Situ, Modification, Activity Assay, Expressing, Marker, Comparison

Journal: Scientific reports

Article Title: Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells.

doi: 10.1038/s41598-019-51778-6

Figure Lengend Snippet: Figure 1. Schematic illustration of the experimental workflow in six steps. B16F1 mouse melanoma cell- derived sEVs were isolated and characterised by Western Blot (WB), atomic force microscopy (AFM) and dynamic light scattering (DLS). Then B16F1 cultures were treated in five different ways, and 72 h supernatants were harvested for sEV isolation. Vesicle samples were then analysed by nanoparticle tracking analysis (NTA) to determine the number of released sEVs, sequencing and LC-MS/MS to describe their miRNome and proteome. Ingenuity Pathway Analysis (IPA) was used to analyse data and predict the functional differences between sEV groups. This in silico predictions were tested in vitro on mesenchymal stem cell (MSC) and melanoma cell cultures and MSC-B16F1 3D co-cultures as well using Ki-67-specific immunocytochemistry, Cell-Clock cell cycle assay, wound healing assay, and 3D hanging drop technology. Abbreviation: n.ctrl-negative control. Figure was created with BioRender.com.

Article Snippet: Purity of MSC cultures was checked by flow cytometry using the Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (R&D Systems) according to the manufacturer’s instructions and a FACSCalibur instrument coupled with CellQuest Pro 6.0 (BD Biosciences).

Techniques: Derivative Assay, Isolation, Western Blot, Microscopy, Sequencing, Liquid Chromatography with Mass Spectroscopy, Functional Assay, In Silico, In Vitro, Immunocytochemistry, Cell Cycle Assay, Wound Healing Assay, Negative Control