Additional file 1 of IRF4 and STAT3 activities are associated with the imbalanced differentiation of T-cells in responses to inhalable particulate matters

Additional file 1: Reagents and Antibodies. Table S1. Primer sequences for qRT-PCR analyses. We designed all the primers using https://sg.idtdna.com/pages/products/custom-dna-rna combine with the NCBI and UCSC websites. Table S2. Chemical characteristics of the PM samples used for functional analyses. Water soluble ions were analyzed by ion chromatography, element measurement by inductivity coupled plasma-mass spectrometry (ICP-MS); and organic carbon (OC) and elemental carbon (EC) fractions by DRI model 2001 carbon analyzer. Figure S1. The average concentration of PM2.5 and PM10 in the three Chinese cities (Beijing, Taiyuan, Shijiazhuang) during the study period. We collected environmental data of 3 weeks preceding the sampling. The actual time period of sampling in the three cities: 10/04/2014–26/04/2014 in Shijiazhuang, Taiyuan: 21/02/2014–01/05/2014 and Beijing: 26/09/2014–16/10/2014. Figure S2. Size distribution and protease activity of the PM sample. A. 20 × microscopic view of PM suspension showing the distribution of PM2.5 and PM10; B. protease activity of the PM samples based on FITC-labeled casein cleavage hydrolysis assay. The protease activities are measured by mean fluorescence level with standard deviation. Figure S3. Expression levels of T-cell related cytokines in mouse serum follow the treatment of PM. Figure S4. Differentiation statuses of T-cell subtypes in mouse lung following the treatment of PM suspension. T-cell differentiation is evaluated by flow cytometry at 18 h, 24 h, 40 h and 72 h after the treatment.

附加文件1：试剂与抗体。
表S1：定量实时逆转录聚合酶链反应（qRT-PCR）分析用引物序列。本研究所有引物均通过https://sg.idtdna.com/pages/products/custom-dna-rna 结合美国国家生物技术信息中心（NCBI）与加州大学圣克鲁兹分校基因组浏览器（UCSC）网站设计完成。
表S2：功能分析所用颗粒物（Particulate Matter，PM）样品的化学特征。其中水溶性离子采用离子色谱法分析，元素组成通过电感耦合等离子体质谱法（inductivity coupled plasma-mass spectrometry，ICP-MS）测定，有机碳（OC）与元素碳（EC）组分则采用DRI Model 2001型碳分析仪进行检测。
图S1：研究周期内中国三座城市（北京、太原、石家庄）PM2.5与PM10的平均浓度。本研究采集了采样前3周的环境监测数据。三座城市的实际采样时段如下：石家庄：2014年4月10日—2014年4月26日；太原：2014年2月21日—2014年5月1日；北京：2014年9月26日—2014年10月16日。
图S2：PM样品的粒径分布与蛋白酶活性。A. PM悬浮液的20倍显微视野，展示PM2.5与PM10的分布情况；B. 基于异硫氰酸荧光素（Fluorescein Isothiocyanate，FITC）标记酪蛋白裂解水解实验的PM样品蛋白酶活性。蛋白酶活性以平均荧光强度及标准差进行表征。
图S3：PM处理后小鼠血清中T细胞相关细胞因子的表达水平。
图S4：PM悬浮液处理后小鼠肺部T细胞亚型的分化状态。于处理后18 h、24 h、40 h及72 h通过流式细胞术评估T细胞分化情况。