1. 利用 Nu-PAGE gel跑出的IP結果不佳2. 整理K-RTA 磷酸化paper投稿至JV後reviewers的意見3. 結果發現CDK9會磷酸化K-RTA 而 S634/S636是可能的位點 但是加入DRB 不會影響K-RTA的分子量 期望能找到合理的解釋4. 大量表達Pin1會造成K-RTA蛋白量降低 而Pin1 inhibitor與 shPin1似乎也會影響K-RTA的表達 量 因此Pin1對於K-RTA stability的影響 需要進一步探討 才能定論(不知道是否因為Pin1 inhibitor或shPin1影響polII的activity呢?)
20110420 Lab meeting-蔡小丸 Read More »
To test the effect of U0126 on EBV Rta-induced EBV reactivation on TW01EREV-2716 1. 初步結果72hr 20uM U0126死了一半的細胞 (WST-1 assay) 2. 感覺Dox 48小時活細胞有隨著U0126濃度的增加而增加 3. Dox 72小時細胞全部懸浮於medium，感覺72小時Dox作用效果遠大於U0126。(這樣可以說U0126是擋不住ER的Function? 也許可以像學姊早上說的降低Dox的量) Next work will 1. Determine the cytotoxicity of U0126 on TW01EREV-2716 by WST-1 assay. 2. Confirm the expression kinetics (0-72hrs) of cell cycle regulators and viral proteins in Dox-treated
04/13 Lab Meeting- 小夏 Read More »
1. 降低細胞數至7.5 x 10^6/ml做sonication的效果仍未達百分百, 將試5 x 10^6/ml的效果 2. 比較TW01TetER(19) Ctrl-24hrs及Dox-24hrs, (1) CTCF bind在H19及SFN promoter的能力似乎相當, 一般PCR及Q-PCR所看到的差距不大 (2) 初步測試Rta也可bind在H19(weak)及SFN promoter上, Q-PCR 確認中
04/06 Lab Meeting 小嬿 Read More »
1. 利用coexpress Flag-K-RTA及Flag-CDK9的lysates 進行CDK9 IP實驗 結果發現K-RTA, S634A/S636A, NLSm均與CDK9有interaction, K-RTA與CDK9的association較S634A/S636A強 2. Pin1 isomerase mutant K34A與K-RTA大量共同表達 K-RTA的molecular weight並沒有改變, 初步推測 Pin1的isomerase的活性並不會影響K-RTA的molecular weight
0330 Lab meeting-蔡小丸 Read More »
HKU-NHRI Joint Symposium Monday, 03-28, 2011Session I Metabolic Disease (Co-chaired by Pak Sham (沈伯松) and Tse-Hua Tan)1. Health Aging Research at HKU (Karen Lam (林小玲))2. Healthy Aging Longitudinal Study in Taiwan (HALST, by Chau Agnes Hsiung, NHRI)3. Adipokines in obesity-related cardio-metabolic complications (Aimin Xu 徐愛民) (goodies: adipokine; baddies: IL6, TNF-a, A-FABP etc)4. Cardiometabolic disease in
心得報告 (KHU-NHRI Joint Symposium) Read More »
(1) 以shLuc、shMGEA5的lentivirus (稀釋倍數1/8) 送入ERKV (2.4×10^5/ml/well/12well) 中，24、48、72小時後，收lysates進行WB確認目標蛋白質的確有減少，B2效果比D2好。shLuc的lysates中，MGEA5的表現隨著時間增加而增加(?)。 (2) 以shLuc、shMGEA5的lentivirus (稀釋倍數1/8) 送入ERKV (2.4×10^5/ml/well/12well) 中，48小時後加Dox，再48小時收lysates進行WB確認，MGEA5減少最多的B2，所引發的lytic reactivation越強烈。加入Dox也會引起MGEA5表現增加(?)。總之，將D2提高至1/4，再加入2個NC(有shRNA但未加Dox)，重做一次。
3/30 Lab Meeting-ingrid Read More »
1. U0126 (MEK1/2 inhibitor) can inhibit EBV Rta induced p21 expression and EBV reactivation in EREV8.2. caspase 3 is involved in the survival activity of DDR1 in OSCC (在OEC-M1 and TW2.6 中knockdown DDR1看到pro form的減少)。3. 用autophage marker LC3B看看autophage是否參予著DDR1調控生長的機制。
03/24 Lab Meeting- 小夏 Read More »
To test Magnetic ChIP method:1. non-specific band was less than agarose method2. By using standard end-point PCR analysis, RNA Pol II is able to bind promoters of GAPDH, H19, and SFN; CTCF can bind H19 promoter and weakly on SFN promoter.3. The problem of sonication efficiency is not dissolved yet.
03/16 Lab Meeting-小嬿 Read More »