Cell migration was determined using a wound healing assay. Cells were plated at a density of 4x105 cells/well in a six-well plate. A pipette tip was used to create a straight wound in each well. Cells were washed twice with PBS (Gibco; Thermo Fisher Scientific, Inc) to remove floating cells and the remaining cells were cultured in DMEM at 37˚C in a humidified incubator with 5% CO2 for 0 and 24 h. Subsequently, cell migration was observed under an inverted phase contrast microscope (magnification, x200; Olympus Corporation). The distance of cell migration was measured by Image ProPlus v6.0 analysis software (Media Cybernetics, Inc.). The relative migration distance = (0 h migration distance -24 h migration distance)/0 h migration distance.
Image proplus v6.0 analysis software
Manufactured by Media Cybernetics
Image ProPlus v6.0 is an image analysis software developed by Media Cybernetics. The software provides tools for capturing, processing, analyzing, and measuring digital images.
Automatically generated - may contain errors
2 protocols using image proplus v6.0 analysis software
1
Wound Healing Assay for Cell Migration
2
Adiponectin-Induced PKCβ1 Translocation
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Immunofluorescence analysis of translocation was conducted as described in our previous studies [26 (link)]. In brief, TG neurons were treated with 100 nM adiponectin for 30 min and then fixed with 4% PFA. After incubation with PBS containing 10% goat serum and 0.2% Triton X-100, neurons were probed with mouse anti-PKCβ1 (1:500, Abcam) overnight at 4 °C and visualized with FITC-conjugated donkey anti-mouse IgG (1:300, ThermoFisher Scientific). The images were captured with a Zeiss LSM510 confocal microscope and analyzed with Image-Pro Plus v6.0 analysis software (Media Cybernetics).
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