Fig 1: Knockdown of SYT13 inhibits cell proliferation. (A) MTT assay was used to evaluate the proliferation of RKO and HCT116 cells with SYT13 knockdown compared with negative control. (B) Celigo cell counting assay was performed to detect the effect of SYT13 knockdown on the proliferation of RKO and HCT116 cells. (C) Images of the colony numbers of RKO and HCT116 cells with or without SYT13 knockdown were captured to examine colony formation ability after 14 days of growth. Data are presented as the mean ± standard deviation (n=3). **P<0.01, ***P<0.001. SYT, synaptotagmin.
Fig 2: Knockdown of SYT13 induces cell apoptosis. (A) Flow cytometry analysis was employed to detect the percentage of apoptotic cells among RKO and HCT116 cells with SYT13 knockdown compared with the negative control. Data are presented as the mean ± standard deviation (n=3). **P<0.01. SYT, synaptotagmin. Knockdown of SYT13 induces cell apoptosis. (B) Cell cycle analysis of RKO and HCT116 cells with SYT13 knockdown and the negative control, and the percentage of cells in the G1, S and G2 phase of the cell cycle. (C) The expression levels of several apoptosis-related proteins, including Bad, Bcl-2, P53 and survivin, were detected by western blot analysis. Data are presented as the mean ± standard deviation (n=3). *P<0.05, **P<0.01, ***P<0.001. SYT, synaptotagmin.
Fig 3: Knockdown of SYT13 inhibits cell migration and invasion. (A) Wound healing assay was used to determine the migration ability of RKO and HCT116 cells with SYT13 knockdown compared with the negative control. Representative images were selected from 3 independent experiments. (B and C) Transwell assay was performed to evaluate the (B) migration and (C) invasion ability of RKO and HCT116 cells with SYT13 knockdown compared with the negative control. Representative images were selected from 3 independent experiments. (D) The expression levels of EMT-related proteins, including N-cadherin, vimentin and Snail, in cells with or without SYT13 knockdown were detected by western blot analysis. Data are presented as the mean ± standard deviation (n=3). *P<0.05, **P<0.01, ***P<0.001. SYT, synaptotagmin; EMT, epithelial-to-mesenchymal transition.
Fig 4: High expression of SYT13 in tumor tissues and construction of SYT13 knockdown cell models. (A) The expression levels of SYT13 in colorectal tumor tissues and paracancerous tissues were detected by immunohistochemical staining (magnification, ×200). (B) The background expression of SYT13 in human colorectal cancer cell lines, including RKO, HCT116, HT-29 and Caco2, was detected by qPCR and western blot analyses, respectively. (C) The infection efficiencies for both cell lines were evaluated by the expression of green fluorescent protein at 72 h post-infection (magnification, ×200). (D and E) The validity and efficiency of SYT13 knockdown were evaluated by (D) RT-qPCR and (E) western blot analysis. Data are presented as the mean ± standard deviation (n=3). *P<0.05, **P<0.01, ***P<0.001. SYT, synaptotagmin; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
Fig 5: Knockdown of SYT13 inhibits tumor growth in a mouse xenograft model. (A) The volume of tumors in shCtrl and shSYT13 groups was measured at 4, 7, 11, 14 and 17 days post-injection. (B) Mean weight of the tumors in the shCtrl and shSYT13 groups. (C) Macroscopic observation of tumors resected from mice in the shCtrl and shSYT13 groups. (D) The tumor sections were subjected to immunohistochemical staining using an antibody against Ki-67 (magnification, ×200). (E and F) The expression of SYT13 in the tumors was detected by (E) quantitative PCR analysis and (F) western blotting. Data are presented as the mean ± standard deviation (n=3). *P<0.05, **P<0.01. SYT, synaptotagmin.
Supplier Page from Abcam for Anti-SYT13 antibody