Matrigel-coated Invasion Assay Protocol

Matrigel-coated invasion assays were performed using a 24-well Transwell chamber system (Corning, NY, USA) as previously described 37 (link). Briefly, 5×10⁴ cells in 400 μL serum-free culture medium were placed into the upper chamber, which was coated with Matrigel (BD, New Jersey, USA). A total of 600 μL medium supplemented with 20% FBS was added to the lower chamber. After incubation for 24 h, cells were fixed in 4% paraformaldehyde and stained with 0.1% crystal violet (Sigma-Aldrich, St. Louis, MO, USA) for 30 min. The stained cells were analyzed.

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Sun T., Peng H., Mao W., Ma L., Liu H., Mai J, & Jiao L. (2021). Autophagy-mediated negative feedback attenuates the oncogenic activity of YAP in pancreatic cancer. International Journal of Biological Sciences, 17(13), 3634-3645.

Publication 2021

Matrigel 24-well Transwell chamber system crystal violet

Assays Cells Cells culture Crystal violet Matrigel Paraformaldehyde Serum

Corresponding Organization : First Affiliated Hospital of Zhengzhou University

Other organizations : Third Affiliated Hospital of Zhengzhou University, Sichuan University, West China Second University Hospital of Sichuan University, West China Hospital of Sichuan University

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Variable analysis

independent variables

Presence or absence of Matrigel coating

dependent variables

Invasion of cells through the Matrigel-coated membrane

control variables

Serum-free culture medium in the upper chamber
600 μL medium supplemented with 20% FBS in the lower chamber
Incubation time of 24 h
Cell density of 5x10^4 cells in the upper chamber

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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