(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 »
No SFN signal in TW01TetEREV#27 cells may be due to GFP antibody-blocking. (Sorry didn’t think of it, I’ll check the probability)
03/02 Lab Meeting- 小夏 Read More »
報告進度(1) TW05TetER：#1-18中，除了#1及#12以外，其餘在IFA中，為FLAG Ab陽性，重複做IFA(點片)以重複確認。(2) 以SK-N-SH cell lysates做OGT Ab的western analysis，覺得沒有認到目標蛋白質，再以Hela cell lysates當positive control及黃老師家的goat二抗再確認。
2/23 Lab Meeting-ingrid Read More »