報告這篇PAPER1. LMP1 increases the release of EGFR into exosomes and that purified exosomes containing LMP1 and EGFR are taken up by epithelial, endothelial, and fibroblast cells, leading to the activation of ERK and PI3K/Akt pathways.2. The ability to detect LMP1 in the exosome pellet harvested from serum of mice carrying the C15 tumor supports a […]
11/17 Lab Meeting- 小夏 Read More »
1. EBV Rta induces growth arrest in the G1 phase of cell cycle in TW01TetER cell lines (the effect is #19＞#7).2. Counting the viable cell by trypan blue exclusion method show that there is no difference between TW01TetER #7 and #19 in the growth rate for the period of 4 days.3. Briefly introduce the NPC
11/3 Lab Meeting- 小夏 Read More »
1. The RNAs of TW02, TW03, TW04, TW05, TW06 are ready. The RNAs of HONE1 and TWO1 will be purified on next week.2. EBV Rta can induce G1 arrest in TW01TetERTA #7, #16 and #19 in my 1st trial (19≧16＞7).3. So far, to test the inhibition effect of U0126 in ERTA-induced EBV reactivation.
10/27 Lab Meeting- 小夏 Read More »
下個月至英國開會Poster內容預演: (1) 簡述 Virapower System。(2) TW01Tet。(3) TW01TetER-7 and -19。(4) TW01-EREVp2089。
08/11 LabMeeting Ingrid Read More »
Teacher蔡: (1) Fig 1C western 太髒, not qualified for publication (要改進!). (2) EAD clone 88 is a good one for IFA (3) Akata strain 比 B95-8 transforms B cell 能力弱很多. (3) Dox-treated ERKV中 knock-downKR (siRNA),用以證實ER是透過KR做事… (4) ACIF是做好EBNA1 IFA之唯一良藥. (5)淑君的Blood paper中述及primary infection可有Z, R, 一直到VCA. (6)葉醫師的differentiation系統為Sphenoidal sinus, 並未有un-differentiated cells. Teacher陳M.-R.: (1) MA 220 is a good antibody
小嬿Progress Report有感 Read More »
以後實驗室quantitate protein band請依以下步驟進行: Plasmid 最好是一起抽的一批, 因實驗室LB/BHI已過使用期限,希望大家每次抽質體時盡量使用新鮮media及LB plate (1-cm厚度最佳),超過一個月以上的media及plate請自行定時清理(請不要當福壽螺到處蔓延留種,謝謝)。 若質體中有SV40-ori, 請以293T為host (DLR/expression etc),其餘它種質體則用293。 Transfection (24~48 h)每次請做p-100/1 ml-lysate的份量(6x p-35/1.2-ml), protein lysate量一次到位,不要老是重複在養細胞/TF效率好像有問題/plasmid不夠…(姑且稱之為小鼻子小眼睛的實驗) 檢測各質體間表現量:(i) 請以wild-type為標準 (所以wild-type可能要多transfect一份),293T為host的取5-ug及10-ug, 293為host的取25-ug及50-ug protein extracts run數片小膠,確定所使用的西方墨漬條件可分出一倍和兩倍間的差異。若是過強,那就請在砍primary/ secondary antibody濃度、incubation時間、ECL強度上三件事情上做調整。(ii)依前步驟所訂出的條件run一片小膠檢查各檢體間是否差異很大,如有二倍以上差異而各步驟又沒有太大疑慮,那表示可能有protein stability/proteosome involved,值得設計其他實驗來分析之.兩倍以下差異者微調input量可繼續往下的實驗。(Plasmid, 細胞用畢的請定時清理,不要當福壽螺到處蔓延留種,謝謝)。 附註定量software有三款,請在Lab Protocol中找到小抄。
首先要謝謝大家的通力合作,希望丟掉/收納一些東西後,大家的手氣會更順! 下週我會把新的list一一印出來,例如protein marker/DNA marker現在都只留一兩根在門上,其餘都已放進新的盒子中.在這段過度時間中,若有東西找不到,請不吝在版上留言.有任何建議或不暸解處,也請提出來討論.另外每天最後離開的人,請幫忙確定一下兩個-20是否都已上鎖!謝謝.