The effect of in vitro heating on the distribution of nuclear matrix polypeptides in HeLa cells |
| |
Authors: | A. M. Martelli R. Bareggi B. M. Riederer R. A. Marugg P. Narducci |
| |
Affiliation: | 4. Biodesign Center for Molecular Design and Biomimetics, Arizona State University, Tempe, AZ 85281, United States |
| |
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 (NH4)2SO4 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. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |