BTF3 promotes proliferation and invasion of PCa cells though modeling stem traits of PCa (a-b) Effect of BTF3 on turmorigenesis in vivo evaluated with xenografts model. PC3 cells with stable expression of Scr/shBTF3 subcutaneously injected into nude mice. Tumors were measured every 3 days using a vernier calliper, and the volume was calculated according to the formula: 1/6 x length x width2. Growth curves of tumors (a), and the average weight of tumor mass in each group (b) were shown. (c) Representative H&E and IHC images of xenograft tumor derived from PC3 Scr/shBTF3 cells. HE and IHC stain were performed to each tumor at harvest time. (d) Cell viability as assessed by MTS assay of indicated PCa cell lines. (e) EdU assays of indicated PCa cell lines transfected with Scr/shBTF3 or Vec/BTF3. Left panel: Representative images of EdU assay. Right panel: Quantitative results of EdU assays from triplicate experiments. (f) The effect of BTF3 on cell migration/invasion evaluated cells by transwell migration and matrigel invasion assays. PCa cell lines were transfected with Scr/shBTF3 or Vec/BTF3 as indicated. Left panel: Representative images of cell migration and invasion. Right panel: Quantitative results of migration and invasion assays from triplicate experiments. In each experiment, cells were counted in five random fields of each filter under a microscope using a 40 × magnification. (g-j) Clonal assay of PC3(G-H) and 22RV1 (I-J) cells with BTF3 ablation or expression. Cells were cultured for two weeks followed by staining with Giemsa and photography. Total number of all clones (g &i) and summary of three types of clones (h &j) presented as representative images (left) and quantification (right). * P < 0.05, **P < 0.01, ***P < 0.001.