July 2011

0720 Lab meeting-蔡小丸

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1. 在293FT細胞中共同表達K-RTA, S634A/S636A, Luc (control protein)及Pin1, 結果發現Pin1降解K-RTA及S634A/S636A的能力相似2. 用一批新抽的K-RTA及S634A/S636A transfection至293FT後發現, K-RTA與S634A/S636A穩定性接近, 推測634/636位點磷酸化與否與其穩定性無關3. 完成oral cancer tissue microarray (BC34011, HN811)之DDR1 IHC染色, 未來將進行DDR1表達量之給分4. 整理DDR1之ligans-collagens(type1-5)在一些oral cancer tissues中觀察到他們的RNA level有增加的情形, 最後我們認為在口腔癌中, collagen type1及type4為可能較重要的DDR1 ligands

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7/13 Lab Meeting-ingrid

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(1) 用IP將KR抓下，用WB(RL2)來確認O-GlcNAc變化程度；結果是可以成功以KR(660) from ICON將KR抓下，但是效率不佳，以致於後來的RL2及PARP1皆無法辨認。接下來會以重收細胞(以EBC Buffer溶解細胞)、增加cell lysate或KR(660)的量來改進IP效率。NOTE：以6% SDS-PAGE分析；KR(Ueda)會認到2條band? (2) 進行Q-PCR檢查送入shOGT的293FT細胞樣本中，MGEA5的mRNA是否有減少，反之亦然。結果發現ACTIN primer會影響，但GAPDH不會影響。經重複實驗，排除off-target作用。(3) TW01TetLuc有4株clones及TW05TetLuc有4株clones進行WB (Flag Ab) 確認為positive。接著進行計數細胞，將生長速率差不多的細胞混合。NOTE：Flag Ab是否有問題?

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EBV Rta-induce cellular senescence in nasopharyngeal cells

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小嬿: (A) 搞定EBV Rta 對 CTCF 在 genome organization 之影響。 (B) 行有餘力 (and a hobby!): Establishment of TW05-EREV Jean: (A) Find out the best differentiation protocol for NP460hTert. (1) reagents: TGFb, calcium … (2) markers: p63, MYC… (3) controls: TW05TetER and TW01TetER #16 cells (4) Ref: 第一篇，第二篇 , 第三篇… (B) 行有餘力: Cloning of TW05-EREV (由小嬿、小夏指導)小夏:Continue

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07/06 Lab Meeting- 小夏

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1. The long-term kinetics (0-6 days) of Rta in Dox-treated TW01TetER The expression of p53, p21, SFN and p27 were increased in Rta induction. Increasing phosphorylation of H2AX was observed on day5 and day6 after Rta induction.2. Compare the expression level of p21, SFN, c-Myc, p53 and p63 in Dox-treated TW01TetER and TW05TetERThe expression of

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