selection of suitable housekeeping genes for real-time quantitative pcr in cd4+ lymphocytes from asthmatics with or without depression选择合适的看家基因的实时定量pcr cd4 +淋巴细胞在哮喘患者有或没有抑郁症.pdfVIP

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selection of suitable housekeeping genes for real-time quantitative pcr in cd4+ lymphocytes from asthmatics with or without depression选择合适的看家基因的实时定量pcr cd4 +淋巴细胞在哮喘患者有或没有抑郁症.pdf

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selectionofsuitablehousekeepinggenesforreal-timequantitativepcrincd4lymphocytesfromasthmaticswithorwithoutdepression选择合适的看家基因的实时定量pcrcd4淋巴细胞在哮喘患者有或没有抑郁症

Selection of Suitable Housekeeping Genes for Real-Time Quantitative PCR in CD4+ Lymphocytes from Asthmatics with or without Depression 1 1 2 1 1 1 Ting Wang , Zong-An Liang , Andrew J. Sandford , Xing-Yu Xiong , Yin-Yin Yang , Yu-Lin Ji *, Jian- Qing He1* 1 Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu, Sichuan Province, People’s Republic of China, 2 The UBC James Hogg Research Centre, Institute for Heart + Lung Health, St. Paul’s Hospital, University of British Columbia, Vancouver, British Columbia, Canada Abstract Objective: No optimal housekeeping genes (HKGs) have been identified for CD4+ T cells from non-depressive asthmatic and depressive asthmatic adults for normalizing quantitative real-time PCR (qPCR) assays. The aim of present study was to select appropriate HKGs for gene expression analysis in purified CD4+ T cells from these asthmatics. Methods: Three groups of subjects (Non-depressive asthmatic, NDA, n = 10, Depressive asthmatic, DA, n = 11, and Healthy control, HC, n = 10 respectively) were studied. qPCR for 9 potential HKGs, namely RNA, 28S ribosomal 1 (RN28S1), ribosomal protein, large, P0 (RPLP0), actin, beta (ACTB), cyclophilin A (PPIA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase 1 (PGK1), beta-2-microglobulin (B2M), glucuronidase, beta (GUSB) and ribosomal protein L13a (RPL13A), was performed. Then the data were analyzed with three different applications namely BestKeeper, geNorm, and NormFinder. Results: The analysis of gene expression data identified B2M and RPLP0 as the most stable reference genes and showed that the level of PPIA was significantly different