Fig. 4.
Detection of Sp1 and Sp3 in nuclear extracts from HeLa and PC3 cells by overlay with specific polyclonal antibodies of a Western blot analysis.
Proteins from HeLa and PC3 nuclear extracts were fractionated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) (10% [wt/vol] for 2 hours at 150 V) and transferred to a PVDF membrane. The Western blot analysis was sequentially overlaid with specific polyclonal antibodies against Sp1, Sp3, and HMGB-1, and protein detection was performed by chemoluminescence. Sp1 as well as all 3 isoforms of Sp3 are present in HeLa and PC3 cell nuclear extracts in comparable amounts (Table 1).