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. 2022 Jul 22;41(35):4145–4158. doi: 10.1038/s41388-022-02411-w

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© The Author(s) 2022

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. 5 — A2780-M cells were transfected with two TRPC3 siRNAs (A) or treated with Pyr3 (B). Addition of external calcium(1.8 mM) led to an increase in fluorescence intensity and representative Ca²⁺ images tracing reflected changes of calcium concentration in the cytoplasm. F/F0: fluorescence normalized to baseline fluorescence. The net change in Ca²⁺ levels was normalized to (F_max − F₀)/F₀. C A2780 cells transfected with TRPC3-overexpressing plasmid or empty plasmid were treated with BAPTA-AM (20 μg/mL, 24 h) or not. Addition of external calcium(1.8 mM) led to an increase in fluorescence intensity and representative Ca²⁺ images tracing reflected changes of calcium concentration in the cytoplasm. F/F0: fluorescence normalized to baseline fluorescence. The net change in Ca²⁺ levels was normalized to (F_max − F₀)/F₀. D A2780 cells transfected with TRPC3-overexpressing plasmid or empty plasmid were treated with BAPTA-AM (20 μg/mL, 24 h) or not. Cellular migration and invasion were detected by transwell assay. Scale bar, 100 μm. E Ovarian cancer cells were orthotopically transplanted into SCID mice, and mice were intravenously treated with Pyr3 (5 mg/kg), BAPTA-AM (5 mg/kg) or DMSO for twice a week from the first week after implantation. Representative bioluminescence images of dissected primary and peritoneal metastatic ovarian cancer at the time of execution. The photon count indicated the tumor burden. F A2780 cells transfected with PLAA siRNA, PLAA siRNA plus BAPTA-AM (20 μg/mL, 24 h) treatment, PLAA siRNA plus TRPC3 siRNA, and negative control, respectively. Addition of external calcium(1.8 mM) led to an increase in fluorescence intensity and representative Ca²⁺ images tracing reflected changes of calcium concentration in the cytoplasm. F/F0: fluorescence normalized to baseline fluorescence. The net change in Ca²⁺ levels was normalized to (F_max − F₀)/F₀. G A2780 cells transfected with PLAA siRNA, PLAA siRNA plus BAPTA-AM (20 μg/mL, 24 h) treatment, and negative control, respectively. Cellular migration and invasion were detected by transwell assay. Scale bar, 100 μm. Data are representative of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.