Research Paper Volume 12, Issue 11 pp 10129—10146

Clusterin exerts a cytoprotective and antioxidant effect in human osteoarthritic cartilage

Figure 6. Effects of CLU silencing on TNFα and ROS levels in cultured OA chondrocytes. (A) Representative immunocytochemical images and evaluation of TNFα+ cytospinned OA-HACs after 72 hours of CLU silencing. (B) Basal mRNA expression of NOX isoforms by RT-PCR in cultured chondrocytes. (C, D) RT-PCR and blot analysis of NOX4 expression in scramble and CLU-silenced OA-HACs. (E, F) Representative images and quantitative measurement of ROS levels (green fluorescence) after 12, 24 and 48 hours of CLU silencing compared with control (scramble) and HACs from fractured patients; original magnification, 100X. Data are expressed as mean values ± SEM of three independent experiments (each using pooled samples from n=3 donors) performed in triplicate. Abbreviations: F-HACs, fractured patient human articular chondrocytes; OA-HACs, OA-Human articular chondrocytes; ADU, arbitrary densitometric unit; F.I., fluorescence intensity. Student’s t-test: *P < 0.05, **P < 0.01 and ***P < 0.001.