Changes in Morphology, Gene Expression and Protein Content in Chondrocytes Cultured on a Random Positioning Machine
Figure 4
Integrity of nuclei counter-stained by Hoechst 33342, distribution of F-Actin, ACTA2 and ACTB gene expression, and Western blot analysis of beta-actin of chondrocytes cultured at 1 g or on the RPM.
A–E: Integrity of nuclei counter-stained by Hoechst 33342 and distribution of F-Actin in human chondrocytes cultured at 1 g or on the RPM for different times (30 min (A), 2 h (B), 4 h (C), 16 h (D), 24 h (E)). After 30 min and 2 h stress fibres were visible in RPM samples (A, B). After 4 h accumulations of F-actin were visible in the outer cellular membrane (C, G). The cells were smaller after 24 h RPM exposure (F). H, I: ACTA2 and ACTB gene expression: Early up-regulation of these mRNAs was found after 0.5 h. J: Western blot analysis of beta-actin: Beta-actin clearly increased after 4 h incubation on the RPM. Data are given as mean ± standard deviation; #P<0.05; *P<0.005.