Purification and biochemical analysis of PCNAs. The wild type and mutant PCNAs were purified and ~1 μg of each protein was analyzed. a
. Purified proteins were resolved in SDS-12 % polyacrylamide gel. Lanes 1, Molecular weight standards; 2, ScPCNA; 3, CaPCNA; and 4, CaPCNA G178S. c. Purified proteins were resolved in 12 % non-denaturing polyacrylamide gel. Lanes 1, CaPCNA; 2, CaPCNA G178S; and 3. ScPCNA. c. Size exclusion chromatograms of wild type (blue) and G178S mutant (red) CaPCNAs with the peak fractionation at ~1.6 ml and ~2.3 ml elution volume, respectively, were shown. d. Glutaraldehyde crosslinked 20 μg of CaPCNA (lane 2) or CaPCNA G178S (lane 3) or ScPCNA G178S (lane 4) proteins were resolved in SDS-12 % polyacrylamide gel. Monomer (~29 kDa) and oligomeric PCNAs (~60-180 kDa) are marked. Lane 1, Molecular weight standards.