Identification of miRNAs which bind to YB-1 protein

To assess the direct association of the nucleic acid binding protein yb-1 with mature microRNAs in breast cancer. To do this we performed immunoprecipitation (IP) of YB-1 protein to pull-down linked RNAs Two breast cancer cell-lines, representative of Luminal A and Basal molecular subtypes were used, MCF7 and MDA-MB-435S. An input control represents the RNA present in the lysate used for the IP, prior to treatment. IPs were performed in triplicates on different cell-lysates with one input control used as a representative of the RNA added to the IP reaction.

本研究旨在探究核酸结合蛋白YB-1与乳腺癌中成熟微小RNA（microRNA）的直接结合关联。为此，我们通过对YB-1蛋白进行免疫沉淀（immunoprecipitation, IP）实验，以富集与其结合的RNA。本研究选用了分别代表管腔A型（Luminal A）与基底型（Basal）分子亚型的两种乳腺癌细胞系：MCF7与MDA-MB-435S。Input对照指代实验处理前，用于IP实验的细胞裂解液中所含的总RNA。本研究针对不同批次的细胞裂解液开展三次重复IP实验，并以一份Input对照作为加入IP反应体系的RNA的代表样本。