Fig. 5

Southwestern analysis of c-Jun-binding activity to the human liver/bone/kidney alkaline phosphatase (L/B/K ALP) 5′-12-O-tetradecanoyl-phorbol-13-acetate (TPA)-response element (TRE) in human periodontal ligament (hPDL) cells upon mechanical stretching. Continuous mechanical stretching was applied to hPDL cell cultures for the indicated time in min prior to harvesting. Whole-cell lysates were prepared and aliquots containing equivalent amounts of protein analyzed by a combined SDS-polyacrylamide gel electrophoresis/protein blotting-renaturation/DNA-binding procedure (southwestern assay). The sequence of the oligonucleotide used as probe is shown (identical to that depicted in Fig. 3). The atypical TRE is highlighted in bold. The molecular mass of the ¹⁴C-methylated aldolase protein as a size marker is shown (M).