Pristimerin Exacerbates Cellular Injury in Conditionally Reprogrammed Patient-Derived Lung Adenocarcinoma Cells by Aggravating Mitochondrial Impairment and Endoplasmic Reticulum Stress through EphB4/CDC42/N-WASP Signaling

Figure 3

Effect of PRIS on cell migration, invasion, and capillary tube formation ability. (a) Chemotactic movement assessed in a Transwell chamber assay with cells seeded in serum-free medium in the upper chamber, with 10% FBS as chemoattractant in the lower chamber. After 48 h incubation, cells on the upper side of the membrane were removed with a cotton swab, insert membrane was stained with Alexa Fluor 488® phalloidin, and the migrated cells were examined using a fluorescence microscope. The number of migrated cells was quantified by performing cell counts of 10 random fields at ×100 magnification (). (b) The upper chambers were coated with 1 : 8 diluted Matrigel Matrix for 1 h at 37°C; cells were seeded in the upper chamber with medium containing 10% FBS in the lower chamber. The following procedures were similar to the migration assay as described above in (a) (). (c) After incubating with conditioned medium from CRLCs for 24 h, HUVECs were grown on Matrigel™ for 18 h under normal growth conditions. Capillary tube formation was observed under an inverted light microscope. Five independent fields were assessed for each well, and the average number of tubes/40x field was determined (). Data are presented as , , .

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