Fig. 4: Analysis of the effect of epithelial cellular adhesion molecule (EPCAM) inhibition and knockdown on the migration of cluster of differentiation (CD)8⁺ T cells in the human leiomyosarcoma cell line TYLMS-1.

a, b TYLMS-1 cells treated with EPCAM inhibitors-produced conditioned media. Tumour necrosis factor-α converting enzyme (TACE) inhibitor (TAPI), and γ-secretase inhibitor (DAPT) were used. In addition to the inhibition of EPCAM signalling by the combination of TAPI and DAPT, partial inhibition of EPCAM signalling was performed with each drug alone. The migration of CD8⁺ T cells was evaluated and compared using a Transwell assay with conditioned medium. c–f TYLMS-1 cells were transfected with small interfering (si)EPCAM or scrambled siRNA-produced conditioned medium. The migration of CD8⁺ T cells was evaluated and compared using a Transwell assay with conditioned medium. a, e The membrane of the Transwell insert was stained, and the number of migrated CD8⁺ T cells was evaluated. Representative microscopy images are shown. b, f Comparison of the numbers of migrated CD8⁺ T cells. For each membrane shown in (a) and (e), the number of stained positive cells was counted in five different fields using a 10× objective lens, and the total number was evaluated. c EPCAM expression in each cell line was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). d EPCAM expression in each cell line was determined by western blotting. Data in (b) and (f) are mean ± standard deviation (SD); n = 3. Two-tailed t-test. *p < 0.05, **p < 0.01.