Generation of cell lines with tetracycline‐regulated autophagy and a role for autophagy in controlling cell size

Hosokawa, Nao; Hara, Yukichi; Mizushima, Noboru

doi:10.1016/j.febslet.2006.04.008

Cited by 194

The DOI matches its Crossref record; 6 of the 6 references it cites were found in a registry or an index.

Generation of cell lines with tetracycline‐regulated autophagy and a role for autophagy in controlling cell size

Hosokawa, Nao, Hara, Yukichi, Mizushima, Noboru·2006·FEBS Letters

DOI 10.1016/j.febslet.2006.04.008

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Abstract

Autophagy is an intracellular bulk degradation system. We established mouse fibroblast lines coupling the Tet‐off system with an Atg5−/− mouse embryonic fibroblast line to artificially regulate autophagic ability. In the presence of doxycycline (Dox), Atg5 expression was completely suppressed and these cells were autophagy‐defective. After removal of Dox, autophagic ability was restored within 6 h. Very low levels of Atg5 could induce an autophagy competent state. We applied this novel system to examine the contribution of autophagy to controlling cell size. Cell size reduction in response to starvation was significantly inhibited in cells unable to undergo autophagy. The generated cell lines will be useful reagents for future mechanistic studies into the regulation and physiologic significance of autophagy.

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“Dendra2 is an improved version of the green-to-red photoswitchable fluorescent protein Dendra, which is derived from the octocoral Dendronephthya sp [346].”
From Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1 (2021) · cites this paper as [346]
Bafilomycin A1 requires ~30 min to increase lysosomal pH; therefore, a preincubation of 30 min is required in case of short autophagy induction times.
“bafilomycin A1 requires ~30 min to increase lysosomal pH [228,346]; therefore, a preincubation of 30 min is required in case of short autophagy induction times.”
From Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1 (2021) · cites this paper as [346]

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Authors: Hosokawa, N., Hara, Y., & Mizushima, N.
Venue: FEBS Letters
Publisher: Wiley
Published: 2006-04-21
Volume: 580
Issue: 11
Pages: 2623-2629
ISSN: 0014-5793
1873-3468
DOI: 10.1016/j.febslet.2006.04.008
Cited by: 194
References: 6
Language: en
Licence: onlinelibrary.wiley.com/termsAndConditions#vor (vor)

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Generation of cell lines with tetracycline‐regulated autophagy and a role for autophagy in controlling cell size

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