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. 2001 Jan 16;98(3):920–925. doi: 10.1073/pnas.021561498

Figure 1.

Figure 1

Hsp27 in HeLa cell crude extracts binds specifically to Rpp20 on His-binding resin. Histidine-tagged Rpp20 was expressed in E. coli (3) and affinity-purified with the use of His⋅Bind resin as described in Materials and Methods. (A) HeLa cell-free extract was added to resin to which Rpp20 was bound, or to resin alone, and proteins that bound specifically to the column were stripped off as described in Materials and Methods. These proteins were separated in a 15% SDS/PAGE gel and were visualized by silver staining (A) or were electrotransferred to nitrocellulose filters and immunoblotted with monoclonal antibody against Hsp27 (StressGen; see B) and Rpp20 (not shown). Leftmost lane: molecular mass markers, molecular masses (in kDa) are indicated at the left of the figure; lane a: 15 ng Hsp27; lane b: 5 ng Hsp27; lane c: 5 ng Rpp20; lane d: proteins from a nuclear extract of non-heat-treated HeLa cells; lane e: proteins from a nuclear extract of heat-shocked HeLa cells.