Journal: Journal of Neuroinflammation

Article Title: Astrocyte elevated gene-1 regulates astrocyte responses to neural injury: implications for reactive astrogliosis and neurodegeneration

doi: 10.1186/1742-2094-9-195

Figure Lengend Snippet: Astrocyte elevated gene-1 (AEG-1) mediates astrocyte migration during wound healing. Human astrocytes were transfected with AEG-1 specific siRNA (siAEG-1) or non-targeting, scrambled siRNA (siCon) by nucleofection and plated for 48 h. Messenger RNA was isolated at 24 h and 120 h post recovery and AEG-1 levels were measured by real-time polymerase chain reaction (PCR) (*** P <0.001, ( A )). In parallel experiments, immunoblotting was performed for AEG-1 at 48 h; β-actin was used as normalizing loading control (** P <0.001, ( B )). Transfected astrocytes were plated to confluence for 48 h into Oris TM migration assay plates and then injured by removal of the cell-seeding stopper. Migrating astrocytes were visualized with Hoechst nuclear stain at various time points. Micrographs are shown for siCon-transfected ( C1 , C3 , C5 , C7 ) and siAEG-1-transfected astrocytes ( C2 , C4 , C6 , C8 ). The number in the lower right corner of the white box (C1-C8) show number of cells in the area of the injury. Separate images from four replicate wells were analyzed to quantify the number of cells present (*** P <0.001, ( D )). Representative data from three individual donors assayed in triplicate is shown.

Article Snippet: The siRNA-transfected astrocytes were plated in 96 well Oris TM cell migration assay plates (Platypus Technologies, Madison, WI, USA) with adherent cell-seeding stopper at a density of 7.5 × 10 4 cells/well and incubated at 37°C for 48 h. Following growth to confluence, the cell-seeding stopper was removed and cells were washed with sterile PBS.

Techniques: Migration, Transfection, Isolation, Real-time Polymerase Chain Reaction, Western Blot, Staining

Journal: eLife

Article Title: Therapeutic downregulation of neuronal PAS domain 2 ( Npas2 ) promotes surgical skin wound healing

doi: 10.7554/eLife.71074

Figure Lengend Snippet:

Article Snippet: A commercially available human dermal fibroblast cell line (CCD-1122Sk, ATCC) (3000 cells/25 µL) was applied to an Oris Pro Cell Migration Assay 384-well plate (PRO384CMA1, Platypus Technologies), which contains a water-soluble biocompatible gel that creates a center cell-free detection zone for cell migration in each well.

Techniques: Cell Migration Assay, Staining, Expressing, Software