Radius 24 well

Cell migration was determined using the Radius™ 24-well from Cell Biolabs (San Diego, CA). In this assay, cells were seeded on Radius cell migration plates and allowed to form monolayers before circular gaps were generated by removing the gels. Cells were then treated with DMSO or different drugs for 24 h, and the migratory gaps were captured at the same magnification using a Zeiss LSM-510 inverted microscope (Zeiss, Germany).

Cell migration was determined using the Radius 24-well from Cell Biolabs (San Diego, CA, USA) according to the manufacturer’s protocol. Cells were seeded and grown to confluence on Radius cell migration plates before circular gaps were generated by removing the gel spot. Cells were then treated with vehicle or WNT5a (600 ng/ml) and incubated at 37 °C for 24 and 48 h. To study the role of Ca²⁺ signaling pathway on migration, cells were treated with KN-93 (2.5 μM/ml) 1 h before treatment with WNT5a, and the phase contrast images of the migratory gaps were acquired using an EVOS XL Core imaging system (Life Technologies, Burlington, Canada). Cell migration into the cell-free area was analyzed using ImageJ analysis software and calculated as the ratio of (surface area on admission-surface area at the time of imaging)/surface area at the time of imaging ×100.

Cell migration was performed using the Radius™ 24-well from Cell Biolabs (San Diego, CA). Briefly, cells were seeded on Radius cell migration plates and allowed to form monolayers. Circular gaps were generated by removing the gels and cells were treated with DMSO or LM11 for 24 hours. To compare differences in the migratory gap, phase-contrast images were captured at the same size using a Zeiss LSM-510 inverted microscope (Zeiss, Germany) and gap closure was analyzed using ImageJ. Cell migration velocity was calculated and statistically analyzed from three independent experiments. Cell invasion was performed using a Matrigel-coated modified Boyden chamber (BD biosciences, San Jose, CA) as described previously [45 (link)–47 (link)]. After incubation for 24 hours, cells on the upside were removed using cotton swabs, and the invading cells on the lower side were fixed and stained with 0.2% crystal violet. Numbers of the invading cells in six randomly selected fields were counted in each experiment using a Zeiss LSM-510 inverted microscope.