| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Research Abstract |
To identify the differentiation factors released in an ischemia cell model using the myoblast-derived cell line, H9c2 cells to the skeletal muscle cells through MyoD, the cell supernatant was separated by ion exchange column, and was examined by MyoD transcriptional activity. The transcriptional activity was undetected in the supernatant. We have carried out proteomic analysis of proteins released from cells during ischemic hypoxia and reoxygenation using H9c2 cells. the cultured supernatant was concentrated more than 10,000-fold by column chromatographies, and the proteins were separated by SDS-PAGE. Proteomic data using nano-LC-Q-Tof-MSMS showed more than 100 proteins specifically released from cells during hypoxic stress compared with control cells. The identified 4 factors were cloned, and introduced in Halo-tag. The cDNAs were transfected in HeLa cells, and partially purified by affinity column. In near future, the factors would be examined in MyoD transcriptional activity.
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