Application of trichostatin A in improving the expression of MHC class Ⅱ molecules in macrophages under microgravity
A technology of macrophages and microgravity, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, material inspection products, etc., to achieve the effect of improving immunity
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Embodiment 1
[0022] C57 / BL6 mouse peritoneal macrophages were cultured under ground gravity and simulated microgravity for 48 hours respectively, and the expression of cell surface molecules CD80, CD86 and MHC class II molecule I-Ab was detected by flow cytometry.
[0023] For the operation method of the above flow cytometry, please refer to D.L. Spector, R.D. Goldman, L.A. Reinwand. Molecular Cloning Experiment Guide Series - Cell Experiment Guide [M]. Huang Peitang, translated. Beijing: Science Publishing Society, 2003: 118.
[0024] The simulated microgravity adopts the simulated microgravity culture system, which was purchased from Shanghai Equl Company, and the model is RCCS-1. The principle of the system simulating microgravity is the same as the rotary culture system invented by NASA, which has been recognized by international counterparts. The usage method of the simulated microgravity culture system is carried out according to the product manual.
[0025] The culture conditions ...
Embodiment 2
[0028] C57 / BL6 mouse peritoneal macrophages were cultured under ground gravity and simulated microgravity conditions for 48 hours, acetylated histone H3, acetylated histone H4, and internal reference protein glyceraldehyde-3-phosphate dehydrogenase The content was detected by Western blot test.
[0029] For the operation method of the above Western blot test, refer to D.L. Spector, R.D. Goldman, L.A. Reinwand. Molecular Cloning Experiment Guide Series - Cell Experiment Guide [M]. Huang Peitang, translated. Beijing: Science Publishing Society, 2003: 639.
[0030] The simulated microgravity adopts the simulated microgravity culture system, which was purchased from Shanghai Equl Company, and the model is RCCS-1. The principle of the system simulating microgravity is the same as the rotary culture system invented by NASA, which has been recognized by international counterparts. The usage method of the simulated microgravity culture system is carried out according to the product ...
Embodiment 3
[0034] C57 / BL6 mouse peritoneal macrophages were cultured under ground gravity and simulated microgravity conditions for 24 hours. Cells for each condition were divided equally into 2 groups. One group was treated with 50nmol / L Trichostatin A for 24 hours (named as Trichostain A group); the other group was left without any treatment and continued to culture for 24 hours (named as control group). The expression of MHC class Ⅱ molecule I-Ab was detected by reverse transcription-fluorescent quantitative PCR
[0035] For the operation method of the above-mentioned reverse transcription-fluorescent quantitative PCR, refer to C.W. Dieffenbacher, G.S. De Foxler. PCR Technology Experiment Guide [M]. Chong Kang, Qu Lijia, translated. Beijing: Science Press, 2006: 131 ..
[0036] The simulated microgravity adopts the simulated microgravity culture system, which was purchased from Shanghai Equl Company, and the model is RCCS-1. The principle of the system simulating microgravity is th...
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