Fig 5.

Characterization of native complexes of class CI, class CII, and class CIII proteins. Tobacco protoplasts were transformed with the indicated combinations of expression plasmids encoding PsHsp18.1-CI (CI), LpHsp17.4-CII (CII), or 3HA-LpHsp16.1-CIII (CIII). After 18 hours at 25°C, native protein extracts were prepared and subjected for separation of sHsp complexes by nondenaturing polyacrylamide gel electrophoresis (A) or by gel filtration chromatography (B), as described in Materials and Methods. Part A shows the results of immunoblot detections on the same blot by sequential cycles of stripping and incubation with antisera specific for class CII (left panel), class CI (right panel), or the HA-tagged Hsp16.1-CIII (middle pannel). In part B, only the elution profiles for Hsp16.1-CIII are shown. Fractions corresponding to the elution of the majority of class CI and class CII proteins are marked by a bracket