Fig. 3: Enhanced migration, invasion, and drug resistance in CD51+ cells.

a The wind-rose plots for the CD51+ and CD51− cells show differences in metastasis. Cells were tracked over time using semi-automated software. The average velocity of dermal CD51+ cells was 0.250 ± 0.017 µm/min, whereas it was 0.163 ± 0.013 µm/min CD51− cells in the DU 145 cell line. Similarly, it was 0.318 ± 0.015 µm/min vs. 0.197 ± 0.011 µm/min in the PC-3 cell line, respectively. b Migration speed was measured and analyzed using a two-tailed t-test. Error bars represent the SD from three independent experiments (***p < 0.001). c Transwell migration assays showed that CD51+ and CD51− cells migrated toward the chemoattractant. Cell migration was determined after 24 h. Scale bar: 200 µm. d Statistical analysis of cell migration. The average was calculated based on three independent experiments. Error bars show the mean ± SD from three technical replicates (two-tailed Student’s t-test, ***p < 0.001). e Wound-healing assay. Phase-contrast images were obtained at 0, 24, and 48 h after scratching the dish. Scale bar: 200 µm. f Statistical analysis of cell migration distance. The average was calculated based on three independent experiments. Error bars show the mean ± SD from three technical replicates (two-tailed Student’s t-test, ***p < 0.001). g H&E staining of tumors that formed in SCID mice injected subcutaneously with CD51+ cells confirmed a malignant phenotype in vivo. Scale bar: 100 µm. h, i Flow cytometric detection of apoptosis and necrosis was performed using annexin V-FITC and propidium iodide (PI). DU 145 (H) and PC-3 (I) cells were divided into three populations: normal viable cells (annexin V and PI negative), early apoptotic cells (annexin V positive and PI negative), and late apoptotic/necrotic cells (annexin V positive and PI positive). Statistical analysis of drug-resistance was performed with a two-tailed t-test. The average was calculated based on three independent experiments. Error bars show the mean ± SD from three technical replicates (two-tailed Student’s t-test, *p < 0.05, **p < 0.01, ***p < 0.001)