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. Author manuscript; available in PMC: 2010 Jul 2.
Published in final edited form as: Arthritis Rheum. 2010 May;62(5):1338–1348. doi: 10.1002/art.27410

Figure 2.

Figure 2

Changes in gene expression and CD44 fragmentation by subculturing chondrocytes in monolayer or alginate beads. A, Dedifferentiation in culture. Total RNA from cultures of primary bovine articular chondrocytes (P0; set at 1.0) or subcultured chondrocytes from P1 to P4 were subjected to real-time reverse transcription–polymerase chain reaction (RT-PCR) analysis. B, Redifferentiation in alginate beads. Chondrocytes from passage 4 were subcultured in monolayer (R0; set at 1.0) or in alginate beads for 1 week (R1) or 2 weeks (R2). Total RNA was isolated and subjected to RT-PCR analysis. C, Western blot analysis of CD44 expression. CD44 in lysates from human knee articular chondrocytes, grown in alginate beads, was detected with BU52 (lane 1) or anti-CD44 cytotail (lane 2). CD44 in lysates from primary bovine articular chondrocytes was detected with anti-CD44 cytotail (lane 3). CD44 in lysates from primary bovine chondrocytes (P0), subcultured chondrocytes (P1–P5), alginate bead–redifferentiated chondrocytes (R1 and R2), or lysates from dedifferentiated bovine articular chondrocytes treated with control small interfering RNA (siRNA; Ctr) or CD44-specific siRNA was detected with anti-CD44 cytotail antibody. Bars represent the mean and SD fold change in mRNA copy number. HAS2 = hyaluronan synthase 2.