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. 2019 Feb 14;38(22):4366–4383. doi: 10.1038/s41388-019-0728-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Effect of SPARC on homing of ovarian cancer (OvCa) cells to omental adipocytes. a In vivo homing of ID8-GFP cells to SP^+/+ and SP^–/– omenta in the presence or absence of prior injection of 5 µg/ml SPARC. Bars represent means ± Standard error of the mean (SEM) of fluorescence intensity of adherent cells to omenta harvested at the indicated time points. *P < 0.05 comparing SP+/+ and SP–/–. **P < 0.05 comparing cells with and without SPARC treatment. ns, non-significant. b Schema of the in vitro homing/chemotaxis of ID8 cells towards SP^+/+ and SP^–/– omental adipocytes. Bars represent means ± SEM of fluorescence intensity of ID8 cells that migrated through trans-wells towards adipocytes. Complete growth media were used as controls for migration (n = 4). *P < 0.05 comparing ID8 migration towards control media, SP+/+, SP–/– adipocytes in absence of SPARC. **P < 0.05 comparing migration of ID8 cells treated with SPARC with the corresponding condition in absence of SPARC treatment. c Schema of the adhesion assay of GFP-OvCa cells over-layed on top of adipocytes for 120 min (left). Bars represent means ± SEM of fluorescence intensity of adherent cells (n = 6/experimental condition). *p < 0.05 comparing ID8 migration towards control media, SP+/+,SP–/– adipocytes in absence of SPARC. **P < 0.05 comparing migration of ID8 cells treated with SPARC with the corresponding condition in absence of SPARC treatment. Photomicrographs of fluorescent adherent cells (top, 5×). d Schema of adipocyte-induced OvCa invasiveness through trans-well inserts towards omental adipocytes in the bottom chamber. Bars represent the means ± SEM of migrated cells counted in five random fields/insert, (n = 3). *P < 0.05 comparing ID8 migration towards control media, SP+/+, SP–/– adipocytes in absence of SPARC. **P < 0.05 comparing migration of ID8 cells treated with SPARC with the corresponding condition in absence of SPARC treatment. Student’s t-test. e Western blots showing the expression of SPARC after overexpression and knockdown in primary human omental adipocytes (hAdi). f Schema of the in vitro homing assay with GFP-labeled human OvCa cell lines. Bars represent means ± SEM of fold change of OvCa cells that migrated through trans-wells towards genetically engineered hAdi compared with cells migrated to control media (without adipocytes) considered as 1. (n = 4/experimental condition. Experiments were repeated three times). *p < 0.05 Student’s t-test with multiple comparisons