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. 2019 Feb 4;10(2):104. doi: 10.1038/s41419-018-1200-y

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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. 6 — a Representative IHC images of normal (left) and tumor (right) TMA sections with BIRC3 staining. For quantification, TMA sections were analyzed by a professional pathologist and scored for 3 (high), 2 (medium), 1 (low), and 0 (negative) of BIRC3 staining. The positive group in the bar figure combined sections with Scores 1–3. b RT-PCR of BIRC3 expression in A2780 and A2780CP cells. Data represent mean ± SD normalized to actin. Results were averaged from three independent experiments, measured in triplicate. Significance of differences was calculated using Student’s t test (****P < 0.0001). c Western blot detection of BIRC3 in A2780 and A2780CP cells. GAPDH serves as loading control. d Western blot detection of BIRC3 in A2780CP cells transduced with virus containing scrambled shRNA (NT) or shRNA against BIRC3. GAPDH serves as loading control. e RT-PCR of BIRC3 expression in A2780CP cells transduced with virus containing scrambled shRNA (NT) or shRNA against BIRC3. Data represent mean ± SD normalized to actin. Results were averaged from three independent experiments, measured in triplicate. Significance of differences was calculated using Student’s t test (***P < 0.001, ****P < 0.0001). f Representative images of A2780CP cell colony formation. Cells were treated with the indicated concentration of cisplatin for 72 h after BIRC3 knockdown, followed by 2 weeks of incubation, and then stained with crystal violet for 30 min. g Western blot detection of BIRC3 in A2780 cells transduced with virus containing BIRC3 overexpression vector or empty PMIG vector as control. GAPDH serves as loading control. h Bar graph showing the effect of BIRC3 overexpression on A2780 cell survival in response to cisplatin treatment (12.5 μM). A2780 cells were transduced with BIRC3-overexpressing virus for 48 h. Cell survival was measured with CCK8 kit 48 h after cisplatin treatment and calculated as described in Fig. 3. Data represent mean ± SD. Results were averaged from three independent experiments, measured in quadruplicate. Significance of differences was calculated using Student’s t test (***P < 0.001). i Flow cytometry analysis showing apoptosis of A2780CP cells transduced with virus containing scrambled shRNA (NT) or shRNA against BIRC3 in response to cisplatin treatment (12.5 μM). Quantification data were averaged from three independent experiments. Apoptosis rate was the sum of the two quadrats on the right in each FACS figure. Data represent mean ± SD. Significance of differences was calculated using Student’s t test (*P < 0.05, **P < 0.01)