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The effect of in vitro heating on the distribution of nuclear matrix polypeptides in HeLa cells

✍ Scribed by A.M. Martelli; R. Bareggi; B.M. Riederer; R.A. Marugg; P. Narducci

Publisher: Elsevier Science
Year: 1994
Tongue: English
Weight: 619 KB
Volume: 18
Category: Article
ISSN: 1065-6995
DOI: 10.1006/cbir.1994.1055

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✦ Synopsis

ABSTRACT

The in situ nuclear matrix was obtained from HeLa cells. After permeabilization with nonionic detergent, the resulting structures were incubated for 1h at 37°C to determine whether or not such an incubation might result in the redistribution of nuclear polypetides which resisted extraction with buffers of high‐ionic strength (1.6 M NaCl or 0.25 M (NH~4~)2SO~4~ as well as DNase I digestion. Using indirect immunofluorescence experiments and monoclonal antibodies we show that heating to 37° C changes the distribution of a 160 kDa protein previously shown to be a component of the inner matrix network. On the other hand, a 125 kDa polypeptide was not affected at all by the incubation. Our results clearly indicate that the inclusion of a 37°C incubation (for example during digestion with DNase I) in the protocol to obtain the in situ nuclear matrix can result in the formation of in vitro artifacts.

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