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hTERT cell line 에 PAH 노출 후 molecular study (cell count, Viability, Copy number variation 등 )

실험계획. hTERT cell line 에 PAH 노출 후 molecular study (cell count, Viability, Copy number variation 등 ) Leukemia cell line - THP-1 cell line 에서 발굴된 Protein { Prelamin -A/C(LMNA), Poly [ADP-ribose] polymerase 1(PARP-1)} 의 mRNA 수준에서 expression study

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hTERT cell line 에 PAH 노출 후 molecular study (cell count, Viability, Copy number variation 등 )

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  1. 실험계획 hTERT cell line에 PAH 노출 후 molecular study (cell count, Viability, Copy number variation등) Leukemia cell line - THP-1 cell line 에서 발굴된 Protein {Prelamin-A/C(LMNA), Poly [ADP-ribose] polymerase 1(PARP-1)}의 mRNA 수준에서 expression study - Western blotting을 이용한 LMNA, PARP-1 Protein 분석: mitochondria fraction, cytosolic fraction - Immunofluorescence stain을 이용하여 PARP-1과 LMNA가 localization되어 있는 위치 확인 3. In vivo Zebrafish model - Western blotting을 이용한 Protein 분석: 시약주문 완료 - Protein (LMNA, PARP-1)의 mRNA 수준에서 expression study SETBP1 mutation in MDS/MPN Patients Heteroplasmic mutation 발견된 sample TA cloning- 시약주문 완료 5. Zebrafish aging study- sequencing완료

  2. Immunofluorescence stain Mitotracker Red CMXRos 1:100 (10nM) 처리 PBS-T(1% BSA 첨가) washing, 5min, 3times PBS:DAPI=10,000:1, 10min PBS-T(1% BSA 첨가) washing, 5min, 2times PBS washing, 5min, 1times

  3. Mitotacker Red Testing 0.01nM-30min 0. 1nM-30min 1nM-30min 10nM-30min

  4. 다음 실험 Plan • Hela cell로 testing • 슬라이드 글라스에 키워 놓음 • -THP-1 cell: Matrigel을 이용하여 고정시킨 후 실험 진행

  5. SETBP1 mutations in MDS/MPN

  6. G2898G/A (Asp868Asp/Asn) Patient No.10 G2898 (Asp868) G2898A (Asp868Asn)

  7. C2903T/C (ser869) Patient No.13

  8. Aging related-zebrafish study

  9. Control region PCR

  10. A339 A339G

  11. C582 C582T

  12. C635 C635T

  13. Mitotracker Red Measurement of early apoptosis Benzopyrene Pentacene 0.6% Normal Fluoranthene DMSO Pyrene

  14. Mitotracker Green Measurement of Mitochondrial Mass Benzopyrene Pentacene Normal Fluoranthene DMSO Pyrene

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