Fig. 2: CST1 promotes GC cell metastasis but not proliferation in vitro.

A, B RT-qPCR and Western blot analysis showing the expression of CST1 in different GC cell lines. Total GAPDH was used as a loading control. C Western blot analysis of HGC-27 and MKN45 stably transfected with CST1 overexpression/knockdown lentiviruses and control lentiviruses. Total GAPDH was used as a loading control. D CCK8 assay analyzed the proliferation of HGC-27-Vector/HGC-27-CST1 and MKN45-shNC/MKN45-sh1-CST1/MKN45-sh2-CST1 stable cell lines. Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by paired t-test. E A colony formation assay. Left panel: representative images, right panel: quantification analysis. Data from independent experiments are presented as the mean ± SD. Statistical was assessed by unpaired t-test, ns means no significance. F Wound healing analysis for assessing migration of HGC-27-Vector/HGC-27-CST1 and MKN45-shNC/MKN45-sh1-CST1/MKN45-sh2-CST1 at 0, 24, and 48 h. Representative images (left panel) and quantification (right panel) are shown as indicated. Data from independent experiments are presented as the mean ± SD. Statistical significance was assessed by an unpaired t-test. ***p < 0.001. Scale bar: 100 μm. G Transwell migration and Matrigel invasion assays were performed to assess migration and invasion ability of CST1-overexpression and knockdown stable cell lines. Representative images (left panel) and quantification (right panel) are shown as indicated. Data from independent experiments are presented as the mean ± SD. Statistical significance was assessed by an unpaired t-test. ***p < 0.001. Scale bar: 100 μm.