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. 2020 Nov 30;10:598684. doi: 10.3389/fonc.2020.598684

Figure 4.

Figure 4

Knockdown of U2AF1 decreased luminal A BC cell dissemination in vivo. Luminal A ER+ MCF-7 cells, transfected with negative control siRNA (siRNA-C) or siRNA targeting U2AF1 (siRNA-U2AF1) were injected in presence of estradiol (E2) ± neutrophils (Neu) into zebrafish transgenic embryos, with green fluorescent blood vessels, and analyzed as described in materials and methods. (A) Migration in vitro (n = 6–12). (B) In vivo dissemination in presence of E2 ± Neu (n = 38–41). Scale bar = 100 µm. (C) Western blot analysis for confirmation of siRNA-U2AF1 transfection and ICAM-1, VCAM-1, and MUC-1 expression. (D) Focal adhesion area (n = 7). Scale bar = 10 µm. (E) Proliferation in vitro (n = 12). Representative images of zebrafish embryos with disseminated MCF-7 cells and immunocytochemistry analysis of vinculin expression are shown. Arrows show disseminated MCF-7 cells and arrowheads show focal adhesions. BV = blood vessels. Data are presented as mean ± SEM. Two-tailed Student’s t-test *P < 0.05, **P < 0.01, ns, not significant. Data are represented of at least two independent experiments.