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 »
(1) Dox誘導型細胞株的選殖TW05TetLuc：#1-6測試中TW05TetER：#2-18的WB有認到ER，重新做IFA以確認表現蛋白質的比例及強弱TW05TetER/gZ：#1-33測試中(2) 以SK-N-SH cell、HelaTet、TW03Tet及TW05Tet lysates做OGT Ab的western analysis，用黃老師家的goat二抗，沒有認到目標蛋白質，重買OGT Ab。(3) 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。
3/16 Lab Meeting-ingrid Read More »
1. 共同表達K-RTA與Pin1後發現, 大量Pin1表達會造成 K-RTA降解2. 初步結果顯示 K-RTA與 Pin1間有交互作用, 但需加入IgG控制組3. 完成silence DDR1對於口腔癌細胞株生長之影響
0309 Lab meeting-蔡小丸 Read More »
1. check Q-PCR primers: p21 and SFN–>Workable!2. ChIP test : a. antibodies: mouse IgG, CTCF and Flag antibody b. primers: H19ICR, SFN, GAPDH(coding region) c. results: (1) the problem of sonication is not dissolved (2) condition of Q-PCR is not well, but standard end-point PCR is OK (3) input is not equal. (one of them
03/02 Lab Meeting-小嬿 Read More »