[bio.dros] [Drosophila] S2 cells (particles/vesicles)

takayuki kuraishi hc12a13 at p.kanazawa-u.ac.jp
Wed Aug 10 21:35:28 EST 2005


Dear Dr. Henrik Fyrst,

I am a graduate student at the Kanazawa University.

I still suspect that your S2 cells were induced to undergo apoptosis.
I have treated S2 cells with Cycloheximide to induce apoptosis
and this treatment causes the presence of a lot of particles, or 
apoptotic bodies.

I would suggest some experiments as follows:
to check condensation of chromatin with Hoechst 33342 staining,
to examine fragmentation of DNA with TUNEL assay or ISNT,
and to look activated Drice with immunocytochemistry instead of Western Blot's.

Reference: J Biol Chem. 279, 48466-76 (2004)

With best regards,

Takayuki

>Hi all,
>
>I'm fairly new to the field of Drosophila S2 cell culture work and would
>greatly appreciate any information that can help me solve this problem. I'm
>doing RNAi experiments and when my protein of interest is knocked down in
>the S2 cells I find the presence of a large number of particles/vesicles
>that almost look like they are moving around. At first these particles are
>seen inside the cells and then later on they are found in the media.
>Does anybody have any idea what these particles might be and what is
>happening to my S2 cell. I have done Western Blot's with antibodies against
>Drice and Dronc and I have also done FACS analysis looking for PS exposure
>by annexin labeling but I haven't found any indication of apoptosis.
>However, treatment of my S2 cells with 20-hydroxyecdysone results in a
>similar scenario with lots of particles. Could it be some sort of immune
>activation?
>
>Sincerely,
>Henrik Fyrst.
>
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-- 
Takayuki Kuraishi 
Grad Sch Med Sci, Kanazawa Univ
Kakuma-machi, Kanazawa
Ishikawa, 920-1192 Japan
Email: hc12a13 at p.kanazawa-u.ac.jp
Phone: +81-76-234-4423
Fax: +81-76-234-4480



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