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【文献标题】Circular RNA-hsa-circ-0000670 promotes gastric cancer progression through the microRNA-384/SIX4 axis

【作者】Pengliang Liu, Shuang Cai, Nuo Li

【作者单位】中国医科大学附属第四医院（the Fourth Affiliated Hospital of China Medical University）

【文献中引用产品】

生物素标记的山羊抗兔抗体

【关键词】Hsa-circ-0000670，MicroRNA-384，Sine oculis-related homeobox 4，Gastric cancer

【DOI】doi.org/10.1016/j.yexcr.2020.112141

【影响因子(IF)】3.21

【出版期刊《Experimental Cell Research》

【产品原文引用】

The tissues were fixed with 4% paraformaldehyde for 24 h, hydrated with 80%, 90%, 100% ethanol and n-butanol, embedded with paraffin, and cut into 5-μm-thick sections. After being heated at 60°C for 1 h, the sections were dewaxed with xylene, dehydrated with gradient ethanol (80%, 90%, 100%) and immersed in 3% H2O2 for 10 min. Antigen retrieval was performed, and the sections were then blocked with 100 μL 5% bovine serum albumin at 37°C for 30 min. Subsequently, the sections were incubated with 100 μL primary rabbit antibodies against SIX4 (1: 2000, ab176713), VEGF (1: 250, ab32152), and CD31 (1: 50, ab28364) overnight at 4°C followed by incubation with a secondary biotin-labeled goat anti-rabbit antibody (HY90046, Shanghai HengYuan Biological Co., Ltd., Shanghai, China) at 37°C for 30 min. Finally, the sections were stained with DAB chromogen (Beijing Bioss Biotechnology Co., Ltd., Beijing, China), and the cell nucleus was stained with hematoxylin for 5 min. The positive cells were stained in brown-yellow. Sections were visualized by Image-Proplus image analysis software (Media Cybernetics, Silver Springs, MD, USA). The target protein expression was evaluated by the proportions of positive cells in total cells detected from 5 randomly selected fields of view on every slide (200-fold magnification). Proportion scores were assigned as follows: positive tumor cells/total tumor cells > 10% positive (+), positive cells < 10% negative (-). 

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