Fig 1: The expression and correlation of SYT7 and TWIST1 in NSCLC tissue determined by IHC and survival analyses and the expression of SYT7 in NSCLC cell lines. a and b, Negative SYT7 protein expression in the normal tissues adjacent to the non-cancerous tissue samples. c, Low expression of SYT7 protein in the adenocarcinoma tissues. d, Low expression of SYT7 protein in the squamous cell carcinoma tissues. e and f, Expression of TWIST1 protein in the tumor tissue shown in c and d, respectively. g, High expression of SYT7 protein in the adenocarcinoma tissues. h, High expression of SYT7 protein in the squamous cell carcinoma tissues. i and j, Expression of TWIST1 protein in the tumor tissue shown in g and h, respectively; a-j were ×400 magnification (scale bar = 50 µm). k, Correlation between TWIST1 and SYT7 protein levels in NSCLC (P < .001, R = 0.403, Pearson's ?2 test). l, Semiquantitative analyses of the SYT7 protein expression in the NSCLC and normal tissues adjacent to the non-cancerous tissue samples (??P < .01, ???P < .001, Mann-Whitney U test). m, The mRNA expression levels of SYT7 in four NSCLC cell lines TWIST1-OE H1975, H125, A549, and H1299. n, Kaplan-Meier survival curves indicated that the protein expression of SYT7 was significantly associated with the OS rate of the clinical LUAD and LUSC patients (Kaplan-Meier method with log-rank test). o, SYT7 was highly expressed in LUAD and LUSC compared with normal lung tissue in the TCGA database (?P < .05, one-way ANOVA test). p, The OS was lower in the patients with a high expression of SYT7 than in those with a low expression of SYT7 in all of the NSCLC patients in the K-M Plotter database (Kaplan-Meier method with log-rank test). q and r, OS was lower in the patients with a high expression of SYT7 than in those with low a expression of SYT7 in LUAD and LUSC patients in the K-M Plotter database (Kaplan-Meier method with log-rank test). The data in i and n are expressed as mean values ± SEM (n = 3).
Fig 2: The selection of TWIST1-downstream genes. a and b, Volcano Plots and Cluster Diagram of differentially expressed genes between TWIST1-OE H1975 cells and control cells. c and d, The HCS experiment and the migration distance ratio of 20 genes in TWIST1-OE H1975 cells. e, The suppression of the migration abilities of A549 cells by SYT7 and CEP85 knockdown. f, Representative photographs of the SYT7 and CEP85 groups and the control group and the comparison of the migration distances between the SYT7 or CEP85 groups with the control group. All of the data are expressed as mean values ± SEM (n = 3).
Fig 3: Association of SYT7 expression with TWIST1 and the regulation function of TWIST1 on SYT7.
Fig 4: Overexpression of SYT7 in vitro and repression of SYT7 in vivo. a, Overexpression efficiency in A549 and H1299 cells confirmed by RT-qPCR and Western blotting. b, MTT assay in the SYT7-OE groups and control groups. c, Colony formation assay. d, Transwell migration assay. e, Invasion assay. f and g, The amount of fluorescence expression in the nude mice model and their lung tissues. h and i, The volume and weight of the tumors formed by the SYT7 knockdown A549 cells and NC A549 cells. All of the data are expressed as mean values ± SEM; ??P < .01, ???P < .001 (Student's t-test).
Fig 5: The repression of SYT7 in regulating metastasis and invasion in vitro. a-d, The expression level of SYT7 mRNA and protein were significantly decreased in the shSYT7 group of the TWIST1-OE H1975, H1299, and A549 cells. e, Knockdown of SYT7 inhibited the metastasis and invasion abilities of the TWIST1-OE H1975 cells. f and g, The metastasis and invasion abilities in the shSYT7 group of A549 and H1299 cells were significantly inhibited in the transwell migration and invasion experiments. All of the data are expressed as mean values ± SEM (n = 3); ?P < .05, ??P < .01, ???P < .001 (Student's t-test).
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