Fig. 6 | Cell Research

Fig. 6

From: Imbalance of the reciprocally inhibitory loop between the ubiquitin-specific protease USP43 and EGFR/PI3K/AKT drives breast carcinogenesis

Fig. 6

USP43 inhibits the proliferation and invasion of breast cancer cells in vitro and suppresses the growth and metastasis of breast cancer in vivo. a MDA-MB-231 cells were infected with lentiviruses carrying control shRNA/vector, USP43 shRNA, USP43, and/or EGFR or MTA3 shRNA for western blotting with antibodies as indicated. b MTS assays for the growth of MDA-MB-231 cells infected with lentiviruses carrying the indicated expression constructs and/or specific shRNAs. Error bars represent the mean ± SD for three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001). c MDA-MB-231 cells infected with lentiviruses carrying the indicated expression constructs and/or specific shRNAs were cultured for 14 days before staining with crystal violet and counting for colony numbers. Error bars represent the mean ± SD for three independent experiments (***p < 0.001). d MDA-MB-231 cells were infected with lentiviruses carrying the indicated expression constructs and/or specific shRNAs for the measurement of the expression of the indicated epithelial/mesenchymal markers by western blotting. The bands were quantified with ImageJ software. The numbers indicate the relative levels of the indicated proteins. e MDA-MB-231 cells were infected with lentiviruses carrying the indicated expression constructs and/or specific shRNA for transwell invasion assays. The invaded cells were stained and counted. The images represent one microscope field in each group. Error bars represent mean ± SD for triplicate experiments (***p < 0.001). Bar, 100 μm. f MDA-MB-231-Luc-D3H2LN cells infected with lentiviruses carrying the indicated expression constructs and/or specific shRNA were inoculated orthotopically onto the abdominal mammary fat pad of 6-week-old female SCID mice. Primary tumor size was measured and metastases were quantified using bioluminescence imaging after 6 weeks of initial implantation. Representative primary tumors and bioluminescent images are shown. Error bars represent mean ± SD for three independent measurements (***p < 0.001). g Representative lung metastasis specimens were sectioned and stained with H&E. Bar, 100 μm. Error bars represent mean ± SD (***p < 0.001)

Back to article page