Figure 3

Immunoblotting of molecules involved in mechano-signal-transduction. PDL cells were seeded on flexible bottom cell culture dishes, strained with averaged 2.5% and followed by westernblot analysis. The numerical expression values were always denoted in relation to unstrained controls (A) The modulation of expression levels of the MAP-kinases p42/44 following strain application were detected with the monoclonal rabbit anti-p42/44 antibody, and the numerical changes of increase and decrease in expression were denoted by arrows in the graph. The phosphorylation levels of Thr²⁰²/Tyr²⁰⁴ of p42/44 were assessed by immunoblotting, using an antibody against phospho- Thr²⁰²/Tyr²⁰⁴. The numerical changes in the activation status of pp42/44 in consequence of strain application were visualised by arrows in the graph. (B) The strain-induced expression levels of the MAP-kinase p38 were detected with a monoclonal rabbit anti-p38 antibody, and the expression changes increase or decrease were marked in the graph by arrows. The strain-induced activation status of phospho-p38 (pp38) was detected by immunoblotting using an antibody against phospho- Thr¹⁸⁰/Tyr¹⁸² of p38, and the modulation of expression levels was notified with numerical values and arrows in the graph. Data of each graph represent the mean of three individual experiments (n = 3), mean +/- SD and means were subjected to the Students T-test. All compared mean values with p < 0.01 were considered as statistical significant and are marked with an asterisk. The depicted western blots exemplify the protein expression changes of one biological replicate.