Fig. 5: Analysis of differentially expressed genes and the effect of synaptosomal-associated protein of 25 kDa (SNAP25) knockdown on cluster of differentiation (CD)8⁺ T cell migration in epithelial cell adhesion molecule (EPCAM)-inhibited and knockdown human leiomyosarcoma cell line TYLMS-1.

a Volcano plot showing differentially expressed genes in TYLMS-1 treated with Tumour necrosis factor-α-converting enzyme (TACE) inhibitor (TAPI) + γ-secretase inhibitor (DAPT) compared to control TYLMS-1 (dimethyl sulfoxide added). Red and blue dots indicate genes with statistically significant differences (fold-change >1.5, p < 0.05). b Volcano plot showing differentially expressed genes in TYLMS-1 with small interfering (si)EPCAM transfection compared to TYLMS-1 with scramble siRNA transfection. Red and blue dots indicate genes with statistically significant differences (fold-change >1.5, p < 0.05). c–f Conditioned media were prepared by transfection of siEPCAM + siSNAP25, siEPCAM, or scramble siRNA into TYLMS-1 cells. The migration of CD8⁺ T cells was evaluated and compared using a Transwell assay with conditioned medium. c The expression of EPCAM and SNAP25 in each cell line was determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). d Expression of EPCAM and SNAP25 in each cell line was determined using western blotting. e The membrane of the Transwell insert was stained, and the number of migrated CD8⁺ T cells was evaluated. Representative microscopic images are presented. f Comparison of the number of migrated CD8⁺ T cells. In each membrane shown in (e), the number of stained positive cells was counted in five different fields using a 10× objective lens, and the total number was evaluated. Data are presented as the mean ± standard deviation (SD), n = 3. Two-trailed t-test. *p < 0.05, **p < 0.01.