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. 2014 May 27;2014:318793. doi: 10.1155/2014/318793

Figure 4.

Figure 4

Silencing of ZNF521 in alginate chondrocyte cultures. (a) Comparison by Q-RT-PCR of the ZNF521 expression in OA chondrocytes (stage III), hematopoietic cell lines (THP1, K562, and IM9), a medulloblastoma cell line (DAOY), and the T/C28a4 chondrocyte cell line. (b) Alginate cultured cells were transduced with lentivirus expressing shRNA and the transgene EGFP; cells were directly visualized for green fluorescence at 20x magnification. (c) Downregulation of ZNF521 mRNA by shRNA lentiviral transduction of chondrocytes in alginate beads. After 3 weeks of culture in alginate beads Q-RT-PCR was performed on the RNA derived from either the control nontarget shRNA or the two shRNA specific for ZNF521. ((d) and (e)) Modulation of the COL2A1 and COL1 ratio by shRNA specific for ZNF521. Chondrocytes from stage III OA were cultured in adherence for 2 passages of trypsinization or in alginate beads with weekly medium changes. At 3 weeks cells were recovered by dissolving the matrix with EDTA and RNA was extracted for Q-RT-PCR and amplified with primers for COL2A1 and COL1.