A. 預計進行 wound healing analysis
(1) OC3 and TW2.6 cells-改變細胞數 (約7-8分滿)+medium中不含collagen
(2) 若有差異 則會再加入 imatinib觀察
B. 預計進行 3D spheroid invasion assay
(1) OC3
(2) 若有差異 則會再加入 imatinib觀察形成球後invasion的情形
目前有的data如下圖所示
C. 預計進行Immunostaining of p-MLC in TW2.6 and OC3cells:
(1) TW2.6 cells 在沒有coating COLI及IV下p-MLC的分佈情形
(2) OC3處理imatinib下p-MLC的分佈情形
Live/Dead staining #1由 maruko 在 四, 07/24/2014 – 16:45 發表。
(AE427 Live/Dead staining Figure ) 看來imatinib處理後 散出去的細胞仍是活的~
#2由 sufang 在 五, 07/25/2014 – 10:37 發表。
Well…
I don't know. 不綠不紅的是算什麼? 感覺上跑出去的細胞大都紅也不綠。然後整體綠光在imatinib這一組是否也下降?
#3由 maruko 在 五, 07/25/2014 – 13:21 發表。
我調整一下對比和強弱
1. 散開大多是綠的啊
2. 加imatinib這組的綠光沒有比較弱(我用眼睛看的) 所以我調了一下對比強弱
3.他其實立體的東西 我不知沒紅沒綠的地方 是focus的問題 或是細胞內的esterase活性降低了 所以綠光淡了 但細胞膜還沒破 所以染不到紅光
#4由 maruko 在 一, 05/05/2014 – 14:43 發表。
AE404
#5由 maruko 在 五, 06/13/2014 – 14:19 發表。
Fig. 6_new
Figure 6. DDR1 contribute to OSCC collective invasion. (a) Effect of imatinib (10 uM) on migration behavior of TW2.6 was determined by time lapse wound healing assay. The images indicate cell migration of the cells for wound closure at the indicated time after scratching (upper panel). The wound healing effect was calculated as the remaining cell-free area compared with the initial area of the original wound (lower panel). (b) 3D spheroid invasion assay were used to observe collective invasion ability of TW2.6 cells. TW2.6 spheroids embedded in collagen I gel were treated with or without imatinib (10 uM) and then observed at the indicated time period. Lower bar graphs showed the relative percentage of collective pushing invaded spheroids in total spheroids was determined by microscopy. (c) DDR1 depleted (upper panel) or imatinib (10 uM) treated TW2.6 cells (lower panel) were immunostained by phospho-S19-myosine light chain (p-MLC, green) and DAPI (blue), and then observed by confocal microscopy.