Fig. 2: JAB1 promotes breast cancer cell proliferation, invasion, and stemness.

A JAB1 knockdown and overexpression efficiency in MDA-MB-231 cells assessed by qRT-PCR and western blotting. B Cell proliferation curves in MDA-MB-231 cells transfected with shSCR, shJAB1, Vector, or FLAG-JAB1. C Example images of EdU assays conducted on MDA-MB-231 cells following transfection with the respective lentiviral Vectors; Scale bar, 100 μm. D Representative images of colony formation assays in MDA-MB-231 cells transfected with the corresponding lentiviral Vectors. E Representative images of cell invasion assays performed with MDA-MB-231 cells using Matrigel Transwell chambers. Expression of epithelial and mesenchymal marker mRNA (F) and protein (G) in MDA-MB-231 cells; J1, JAB1. H Lung metastasis was observed by small animal imaging after tail vein injection of MDA-MB-231 cells. I Images of mouse lung nodules and statistical analysis. J H&E staining of mouse lung tissue. K Representative images of mammosphere formation assays in MDA-MB-231 cells show both the number (left panel, scale bar: 200 μm) and diameter (right panel, scale bar: 100 μm) of spheres after 15 days of culture. Expression of stemness markers mRNA (L) and protein (M) in MDA-MB-231 cells; J1, JAB1. N JAB1 knockdown suppressed the stemness of breast cancer in vivo. MDA-MB-231 cells stably transfected with shSCR and shJAB1 were injected into the fourth pair of mammary fat pads of NOD/SCID female mice (n = 6 per group). O Tumor growth curves of NOD/SCID mice injected with shSCR or shJAB1 MDA-MB-231 cells at different cell doses. J1, JAB1; The error bars indicate the mean ± SD derived from three separate experiments; two-tailed unpaired t-test; *p < 0.05, **p < 0.01, ***p < 0.001.