Fig. 2: OTUDin3 inhibits lung cancer cell growth, migration and invasion, and induces apoptosis.

A Cell proliferation assays in H1299 or H460 cells treated with increasing concentrations of OTUDin3 for 72 h. Viability of cells was determined by the CCK-8 kit. IC₅₀ was analyzed by nonlinear regression (curve fit) using GraphPad Prism V7.0 software. B Representative images of colony formation assays in H1299 cells treated with increasing concentrations of OTUDin3 for 2 weeks after seeding of 1000 cells in six-well plate. Scale bar: 1 cm. C H1299 cells were wounded and then cultured in medium containing increasing concentrations of OTUDin3 for 24 h. Representative images of wound-healing assay were showed. Scale bar, 100 μm. D Representative images of invasion assays of H1299 cells treated with increasing concentrations of OTUDin3. Scale bar: 100 μm. E, F H1299 cells were treated with increasing concentrations of OTUDin3 for 24 h. The apoptotic cell population including early and late apoptosis cells was quantified using an apoptotic kit and analyzed by flow cytometry. The data were analyzed by FlowJo_V10.8.1 software (Treestar, Ashland, OR). The column chart (F) shows the quantification of apoptotic rate of H1299 cells after OTUDin3 treatment. G Western blotting analyses of cell lysates were performed with antibodies against Cleaved-PARP, Cleaved-Caspase-3, PARP, Caspase-3, OTUD3, and GAPDH, as indicated. A representative image of three independent experiments is shown. All the data shown are mean ± SD. n = 3 or 4 independent experiments. Two-tailed unpaired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001, ns = no significance, vs control group.