Matrigel mix

For Transwell assays, according to the manufacturer’s protocol, GC cells were seeded in upper chambers with 200ul of serum-free medium. The transwell chamber (Corning, USA) was paved with matrigel mix (BD Biosciences, USA) for invasion assays and paved without matrigel mix for migration assays. The bottom chamber was filled with medium and 10% FBS as a gastric cancer cell chemoattractant. After incubation for 24 h, the upper chambers were fixed and then stained by crystal violet (Kaigen, China) for 15 min. For visualization, the cell lines were photographed and counted in different five fields. These experiments were repeated for three times.

According to the manufacturer's instructions, we inoculated processed YY8103 and Hep3B cells into 200 μl serum-free DMEM medium in the upper chamber. The transwell room (Corning, USA) accepted the paving process, used Matrigel mix (BD Biosciences, USA) for the invasion test process, and did not use the Matrigel mix for the migration test. DMEM medium and 10% FBS are introduced into the bottom chamber to become HCC cytochemical attractants. When the 48-hour incubation process is completed, the upper chamber is fixed and then stained with crystal violet (Kagan, China) for 15 minutes. For the visualization process, the cell line received the photo and counting process in three fields.

We used the transwell chamber (Corning, NY, USA) to conduct the assays of cell migration and invasion. The transwell chamber was coated with the matrigel mix (BD Biosciences, San Jose, CA, USA) for invasion assay, and it could also be conducted for migration assay with matrigel mix coat.
A549 and H1299 cells at the logarithmic phase were digested and diluted to 1×10⁵/ml with the invasion medium of IMDM + 0.1% BSA without serum. About 100 µl cells were added on top of the transwell membrane in the upper chamber, and 600 µl of IMDM + 10% FBS + the related drugs were added to the lower chamber in a 24-well plate. After the drug treatment for 48 h, we used cotton swabs to scrape the cells settled on the upper surfaces of the transwell chambers and fixed the cells settled on the lower surfaces. The remaining cells were stained by DAPI. Next, we observed the number of cells in the transwell chambers under a fluorescent inverted microscope and took a photo later.

By complying with the producer's guidelines, MCF-7 and ZR75-1 cells received seeding in the upper chambers with 200 μl of serum-free RPMI 1640 medium. The transwell chamber (Corning, NY, USA) received paving process with Matrigel mix (BD Biosciences, San Jose, CA, USA) for invasion testing process and with no use of Matrigel mix for migration tests. RPMI 1640 medium and 10% FBS received the introduction inside the bottom chamber to be a BC cell chemoattractant. When the 24 h incubating process was achieved, the upper chambers received the fixing, followed by staining based on crystal violet (Kaigen, Nanjing, China) for 15 min. For the visualizing procedure, the cell lines received the photograph and count procedures inside 5 fields.