LGR5 expression relative intensity in porcine cochleae

LGR5 expression relative intensity in porcine cochleae from E80, P0, P60, and P120 is collected at six different frequencies of 40 Hz, 100 Hz, 500 Hz, 1000 Hz, 10000 Hz, and 20000 Hz. To quantify the LGR5 expression, the LGR5 intensity of 10 individual cells from each subset of supporting cells including greater epithelial ridge/inner sulcus cells (GER/ISC), inner border cells (IBC), inner phalangeal cells (IPHC), inner and outer pillar cells (IPC, OPC),1st and 2nd Deiters’ cells (DC12), 3rd Deiters’ cells (DC3), and Hensen cells (HS) on surface view was measured via ImageJ and divided by the background intensity at 6 different frequencies. We divided the LGR5 intensity by the background to normalize the signal strength at different depths and positions, under the assumption that the background intensity i.e., autofluorescence should be equal in all cell locations in the organ of Cortis for an individual sample. A total of 16 cochleae (LGR5-H2B-GFP) were subject to quantification: 4 cochleae were taken from E80 pigs, 4 cochleae were taken from P0 pigs, 4 cochleae were taken from two P60 pigs, and 4 from P120 pigs.本数据集采集了胚胎第80天（E80）、出生后第0天（P0）、第60天（P60）及第120天（P120）的猪耳蜗中LGR5的相对表达强度，检测覆盖6种指定频率：40 Hz、100 Hz、500 Hz、1000 Hz、10000 Hz及20000 Hz。 为定量LGR5的表达水平，研究人员通过ImageJ软件测量了表面观下各支持细胞亚群的10个单个细胞的LGR5荧光强度，涉及的支持细胞亚群包括：上皮嵴/内沟细胞（greater epithelial ridge/inner sulcus cells, GER/ISC）、内边界细胞（inner border cells, IBC）、内指细胞（inner phalangeal cells, IPHC）、内柱细胞与外柱细胞（inner and outer pillar cells, IPC、OPC）、第1、2代戴特细胞（1st and 2nd Deiters’ cells, DC12）、第3代戴特细胞（3rd Deiters’ cells, DC3）以及亨森细胞（Hensen cells, HS）。随后将各细胞的LGR5强度除以对应频率下的背景强度，对不同成像深度与位置的信号强度进行归一化；本次归一化的前提假设为：单个样本的柯蒂氏器（organ of Corti）中所有细胞位置的背景强度（即自发荧光）均保持一致。 本研究共纳入16个LGR5-H2B-GFP转基因耳蜗样本进行定量分析：其中4个取自E80胎猪，4个取自P0新生仔猪，4个取自2头P60龄猪，剩余4个取自P120龄猪。