Differentiation and Activation of Dendritic Cells

BMDCs were obtained as the method described with some modifications43 44 (link). On incubation day 3 and 6, fresh medium containing granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4 was added to the plate. On incubation day 7, TJ-M2010-2 was added (0 μM, 10 μM and 40 μM) for 1 h, after which LPS, (1 μg/ml45 (link)46 (link), Sigma, St. Louis, MO, USA) was added for an additional 48 h. Finally, non-adherent cells were collected and stained with FITC-conjugated anti-CD80, PE-conjugated anti-MHC-II, PE-Cy5-conjugated anti-CD86 and APC-conjugated anti-CD11c (all from eBioscience, San Diego, CA, USA) for analysis by flow cytometry (FCM).

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Zhang L.M., Liu J.H., Xue C.B., Li M.Q., Xing S., Zhang X., He W.T., Jiang F.C., Lu X, & Zhou P. (2016). Pharmacological inhibition of MyD88 homodimerization counteracts renal ischemia reperfusion-induced progressive renal injury in vivo and in vitro. Scientific Reports, 6, 26954.

Publication 2016

LPS FITC-conjugated anti-CD80 PE-conjugated anti-MHC-II PE-Cy5-conjugated anti-CD86 APC-conjugated anti-CD11c

Anti cd80 Cells Fitc Flow cytometry Granulocyte macrophage colony stimulating factor

Corresponding Organization :

Other organizations : Huazhong University of Science and Technology, Tongji Hospital, Zhongnan Hospital of Wuhan University, Wuhan University, Taian City Central Hospital, Ministry of Education of the People's Republic of China

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Variable analysis

independent variables

TJ-M2010-2 concentration (0 μM, 10 μM, 40 μM)

dependent variables

CD80 expression
MHC-II expression
CD86 expression
CD11c expression

control variables

GM-CSF and IL-4 in the medium on incubation day 3 and 6
LPS (1 μg/ml) for 48 h

controls

Positive control: LPS stimulation
Negative control: 0 μM TJ-M2010-2

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