報告進度
(1) TW03Tet有10株細胞株，TW05Tet有8株細胞株，已凍管且收好cell lysates等待Western analysis確認。
(2) 目前要進行的實驗如下
a. 將不同稀釋倍數(0、1、1/2、1/4、1/8、1/16)的pAS-EGFP lentivirus送入293中，48小時後，計數細胞存活及死亡比率，同時跑FLOW。(這次infection時，不從細胞上移除含有polybrene的medium)
b. 以shLuc、shOGT、shMGEA5的lentivirus送入ERKV中，24、48、72小時後，收lysates進行WB確認目標蛋白質是否有減少。
c. 將OGT Ab的條件抓好。
Dec 30, 2010
Dec 27, 2010
12/22 Lab Meeting-ingrid
報告Hart新作：
(1) 以chemoenzymatic assay及抗體確認histone有O-GlcNAcylation
(2) 以LC MS/MS定義出histone上有被O-GlcNAc修飾的位點：Ser47 of H4、Ser36 of H2B和Thr101 of H2A
(3) Heat shock(45℃，1hr)及後續的recovery的時候，發現histone的O-GlcNAc程度逐漸提升，同時OGT活性也有緩慢上升
(4) 以MNase chromatin-sensitivity assays檢測發現Heat shock及後續的recovery時候，Chromatin(染色質)會condense
(5) 過量表現OGT會比過量表現GFP的細胞的染色質更加condense
(1) 以chemoenzymatic assay及抗體確認histone有O-GlcNAcylation
(2) 以LC MS/MS定義出histone上有被O-GlcNAc修飾的位點：Ser47 of H4、Ser36 of H2B和Thr101 of H2A
(3) Heat shock(45℃，1hr)及後續的recovery的時候，發現histone的O-GlcNAc程度逐漸提升，同時OGT活性也有緩慢上升
(4) 以MNase chromatin-sensitivity assays檢測發現Heat shock及後續的recovery時候，Chromatin(染色質)會condense
(5) 過量表現OGT會比過量表現GFP的細胞的染色質更加condense
12/22 Lab Meeting-蔡小丸
1. 成功移除OEC-M1內的mycoplasma
2. 比較多株口腔癌細胞DDR1 autophosphorylation的情形 初步結果顯示台灣地區口腔癌細胞株的DDR1 tyrosine phosphorylation的程度較高
3. 一系列K-RTA vs. CDK1,2,6,9的in vitro kinase assay 仍須再試條件 讓結果更明顯 希望會成功!!!
2. 比較多株口腔癌細胞DDR1 autophosphorylation的情形 初步結果顯示台灣地區口腔癌細胞株的DDR1 tyrosine phosphorylation的程度較高
3. 一系列K-RTA vs. CDK1,2,6,9的in vitro kinase assay 仍須再試條件 讓結果更明顯 希望會成功!!!
Dec 21, 2010
12/15 Lab Meeting-小嬿
報告進度:
1. TW01EREV_S無法用SAHA induction成功-->放棄S
2. 所建立的Akata-p2089, 用hIgG沒有辦法induction(by Q-PCR), 在induce 96hrs情況下, 看不到Rta, Zta及EAD表現, 但有少量的gp350/220. 根據之前研究, 可續偵測LMP2A的存在與否.
3. Akata-EGFP/EBV在hIgG induce 48hrs可到達plateau (in sup, by Q-PCR).
4. 目前所建立的35個TW01EREV中, EBV Rta表達量高者似乎無法induction, 僅有TW01EREV-26可用NaB產出較高量的病毒; 在EBV Rta表達中量者中, 挑到1個Dox與 NaB效果相當-TW01EREV-27. 但其induction rate, viral particles等細項仍須confirm.
5. 搜尋其他研究者的ChIP sonication 條件.
1. TW01EREV_S無法用SAHA induction成功-->放棄S
2. 所建立的Akata-p2089, 用hIgG沒有辦法induction(by Q-PCR), 在induce 96hrs情況下, 看不到Rta, Zta及EAD表現, 但有少量的gp350/220. 根據之前研究, 可續偵測LMP2A的存在與否.
3. Akata-EGFP/EBV在hIgG induce 48hrs可到達plateau (in sup, by Q-PCR).
4. 目前所建立的35個TW01EREV中, EBV Rta表達量高者似乎無法induction, 僅有TW01EREV-26可用NaB產出較高量的病毒; 在EBV Rta表達中量者中, 挑到1個Dox與 NaB效果相當-TW01EREV-27. 但其induction rate, viral particles等細項仍須confirm.
5. 搜尋其他研究者的ChIP sonication 條件.
Nov 24, 2010
11/24 Lab Meeting-蔡小丸
1. 除了OEC-M1以外 其他的口腔癌細胞株都沒有mycoplasma的污染
2. 多株口腔癌細胞株DDR1表達量之比較
3. shDDR1及Glivec對於非台灣地區 口腔癌細胞的影響
4. 持續測試OSCCs之DDR1 activity (phospho-Tyrosine status)
2. 多株口腔癌細胞株DDR1表達量之比較
3. shDDR1及Glivec對於非台灣地區 口腔癌細胞的影響
4. 持續測試OSCCs之DDR1 activity (phospho-Tyrosine status)
Nov 20, 2010
Nov 18, 2010
11/17 Lab Meeting 小嬿
1. Establishment of Akata-p2089, TW01EREV-Akata-EGFP cell lines.
2. To test the sonication condition of ChIP.
3. To evaluate the expression level of X protein in 293- and TW01-series cell lines.
2. To test the sonication condition of ChIP.
3. To evaluate the expression level of X protein in 293- and TW01-series cell lines.
Nov 17, 2010
11/17 Lab Meeting- 小夏
報告這篇PAPER
1. 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 physiological role of LMP1-containing exosomes in NPC.
3. EBV miRNAs have been detected in NPC exosomes.
4. The effects of EBV and LMP1 on exosomes are likely to be important factors in EBV infection through which a small number of cells expressing LMP1 could both modulate the tumor microenvironment and also potentially impact the infected host.
1. 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 physiological role of LMP1-containing exosomes in NPC.
3. EBV miRNAs have been detected in NPC exosomes.
4. The effects of EBV and LMP1 on exosomes are likely to be important factors in EBV infection through which a small number of cells expressing LMP1 could both modulate the tumor microenvironment and also potentially impact the infected host.
Nov 12, 2010
Nov 11, 2010
11/10 Lab Meeting- ingrid
1. 想要測試Dox對TW01Tet的生長是否有所影響，結果失敗。下次要收短時間測試(如1、2、3天)。
2. 篩選TW03Tet及TW05Tet細胞株。TW03Tet：24株裡有19株長起來；TW05Tet：24株裡有12株長起來。目前皆放在6 well內等待Western blot測試TetR。
3. OGT抗體無法辨認抗原，已重新購買。另外要解決internal control在每個sample中的量皆不相同的問題。
4. 將shMGEA5送入ERKV細胞中，”似乎”可以使發紅光的細胞比例上升，要再進一步詳細確認。
2. 篩選TW03Tet及TW05Tet細胞株。TW03Tet：24株裡有19株長起來；TW05Tet：24株裡有12株長起來。目前皆放在6 well內等待Western blot測試TetR。
3. OGT抗體無法辨認抗原，已重新購買。另外要解決internal control在每個sample中的量皆不相同的問題。
4. 將shMGEA5送入ERKV細胞中，”似乎”可以使發紅光的細胞比例上升，要再進一步詳細確認。
Nov 7, 2010
33 Lines
33 Lines
由 sufang 在 日, 11/07/2010 - 00:00 發表 Pre-published 33 Lines Cell Lines Dan Robinson PAPA PTK profiling RAGE
由 sufang 在 日, 11/07/2010 - 00:00 發表 Pre-published 33 Lines Cell Lines Dan Robinson PAPA PTK profiling RAGE
| Cell Designation | Tissue 1 | Tissue 2 | Morphology | |
| SW-13 | Adrenal gland | cortex, primary small cell carcinoma | Epithelial | |
| T24 | Bladder | transitional cell carcinoma | Epithelial | |
| SW-1353 | Bone | chondrosarcoma | Fibroblast | |
| Saos-2 | Bone | Osteosarcoma | Epithelial | |
| K-562 | Bone Marrow | Chronic myelogenous leukemia | Lymphoblast | |
| Daoy | Brain | desmoplastic cerebellar medulloblastoma | polygonal | |
| SK-N-SH | Brain | Meta: bone marrow neuroblastoma | Epithelial | |
| SK-BR-3 | Breast | Meta: pleural effusion adenocarcinoma | Epithelial | |
| SW620 | Colon | Meta: lymph node colorectal adenocarcinoma | Epithelial | |
| HT-1080 | Connective tissue | Fibrosarcoma | ||
| U251 | Glioma | AdherentFibroblast | ||
| Caki1 | Kidney | Meta: skin clear cell carcinoma | Epithelial | |
| MOLT-4 | Leukemia | ALL | T Lymphoblast | ITK |
| Jurkat | Leukemia | ALL | T Lymphoblast | ITK |
| CEM | Leukemia | ALL | T Lymphoblast | ITK |
| MEG-01 | Leukemia | CML | megakaryoblast, Lymphoblast | ABL1 |
| HL-60 | Leukemia | promyeloblas, acute promyelocytic leukemia | myeloblastic | |
| HepG2 | Liver | hepatocellular carcinoma | Epithelial | |
| A549 | Lung | Carcinoma | Epithelial | |
| NCI-H69 | Lung | Sm Cell Lung Cancer | floating aggregates | |
| Ramos | Lymphoma | Burkitt's lymphoma, EBV - | Lymphoblast | |
| U-937 | Lymphoma | Histiocytic, EBV - | Monocyte | |
| RD | Muscle | rhabdomyosarcoma | spindle cells and large multinucleated cells | |
| SK-OV-3 | Ovary | Meta: ascites adenocarcinoma | Epithelial | |
| Capan-1 | Pancreas | Meta: liver adenocarcinoma | Epithelial | |
| LNCaP | Prostate | Meta: left supraclavicular lymph node carcinoma | Epithelial | |
| SW837 | Rectum | adenocarcinoma | Epithelial | |
| A-431 | Skin | Epidermoid carcinoma | Epithelial | EGFR |
| SK-MEL-28 | Skin | malignant melanoma | polygonal | |
| AGS | Stomach | gastric adenocarcinoma | Epithelial | |
| SW-982 | Synovium | synovial sarcoma | Mixed | |
| HEC-1-A | Uterus | endometrium; adenocarcinoma | Epithelial | |
| HUVEC | Vein | Umbilical Vein Endothelial Cells | BMX, KDR, TEK | |
| Path at iMac: SORTED/2010/2010_PAPA_33Lines.xlsx | ||||
Nov 4, 2010
11/3 Lab Meeting- 小夏
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 cell lines established from Dr. Chin-Tarng Lin and published in 1990 and 1993.
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 cell lines established from Dr. Chin-Tarng Lin and published in 1990 and 1993.
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