In vitro transcription studies used in a proof of concept whole transcriptome model predition study - HepaRG cells (3 of 4)

HepaRG cells were exposed in triplicate to three agrichemicals for 24hrs at 8 concentrations and a DMSO vehicle control (0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM plus DMSO vehicle controls). While a common set of DMSO controls was used, these CEL files were RMA normalized independently with each of the chemical treated groups. Gene expression was measured on an Affymetrix GeneTitan system. The compounds used were fenbuconazole (a.k.a FENB, CAS # 114369-43-6) a triazole fungicide, imazalil (a.k.a. IMAZ, CAS # 35554-44-0), an azole pesticide, and 2,4-dichlorophenoxyacetic acid (a.k.a. 2,4-D or 2-4-D in file names, CAS # 94-75-7), a chlorophenoxy herbicide. 24hr in vitro exposures in triplicate at 9 concentrations - 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM plus DMSO vehicle controls

将HepaRG细胞以三份重复的方式，分别暴露于三种农用化学品中，每个化学品设置8个浓度梯度及二甲基亚砜（DMSO）溶剂对照组，处理时长为24小时，浓度梯度为0.001、0.003、0.01、0.03、0.1、0.3、1、3、10 μM，同步配套DMSO溶剂对照。尽管所有组别共用一套统一的DMSO溶剂对照，但所有经化学品处理的组对应的CEL文件均采用RMA（稳健多阵列平均法）独立进行归一化处理。基因表达谱通过Affymetrix GeneTitan系统完成检测。本次实验所用化合物如下：糠菌唑（fenbuconazole，简称FENB，CAS登记号114369-43-6），为三唑类杀菌剂；咪鲜胺（imazalil，简称IMAZ，CAS登记号35554-44-0），为唑类农药；以及2,4-二氯苯氧乙酸（2,4-dichlorophenoxyacetic acid，文件名中简称2,4-D或2-4-D，CAS登记号94-75-7），为氯苯氧基类除草剂。本实验采用三份重复，设置9个浓度梯度（0.001、0.003、0.01、0.03、0.1、0.3、1、3、10 μM）外加DMSO溶剂对照，开展24小时体外暴露处理。