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. 2021 Aug 4;12:713332. doi: 10.3389/fphar.2021.713332

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Copyright © 2021 Kato, Pimenta, Almeida, Zambelli, Santos and Sampaio.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of the CTX on migration assay induced by MCF-7-Conditioned medium. (A) Wound healing assay performed in CTX-pretreated t.End.1 cells in the presence of RPMI medium containing 10% FBS (control) or in the presence of MCF-7-conditioned medium (chemotactic stimulation) for a period of 24 h. After this time, cells were fixed by the Rosenfeld panchromic method. A total of five random fields per coverslip were counted under a brightfield microscope (Standard 25; Carl Zeiss, Germany) using a ×10 objective. The results are expressed as Number of Migrated Cells and represent the mean ± s.e.m. The data are presented from three distinct experiments run in quadruplicate of each group. *p < 0.001 compared to RPMI control group. **p < 0.001 compared to RPMI control group. #p < 0.001 compared to MCF-7-CM control group. The images represented in (B) in Panels (a–d) were obtained in a ×5 objective. Panel (a) represents control cells incubated in RPMI 1640 medium with 10% FBS; Panel (b) represents cells treated with CTX and incubated in RPMI 1640 medium with 10% FBS; Panel (c) represents control cells incubated in MCF-7-CM and Panel (d) represents CTX treated cells and incubated in MCF-7-CM. For the Transwell assay, in (C) the number of migrating cells was determined by counting in five random fields per membrane using light microscopy. The results are expressed as Migrant Cells and represent the mean ± s.e.m. *p < 0.001 compared to RPMI control group. #p < 0.01 compared to MCF-7-CM control group. (D) The panels are representative of t.End.1 cell chemotaxis: (a) control in response to RPMI 1640 medium with 2% FBS; (b) CTX in response to RPMI 1640 medium with 2% FBS; (c) control in response to MCF-7-CM; (d) CTX in response to conditioned medium from MCF-7 tumor cells. The images were collected under an Olympus BX 51 microscope using the Image-Pro Plus 5.1 program, using a ×40 objective. The data are presented from three distinct experiments.