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. 2016 May 5;11(5):e0154951. doi: 10.1371/journal.pone.0154951

Fig 1. Overview of experimental design.

Fig 1

(A) Workflow scheme for analysis of the effect of cyclic equibiaxial tensile strain (CTS) on HDAC4 relocation in chondrocytes. After isolated and then cultured for 6 days, the passage chondrocytes were transfected with GFP-HDAC4. At 48 h post-transfection, the transfected cells were submitted to CTS for 3 hours. (B) The chondrocytes at Area A were subjected to 6% equibiaxial CTS. (C) Green fluorescent protein (GFP) was captured by fluorescence microscope to validate the transfection efficiency of GFP-HDAC4 in chondrocytes. Nuclei were stained by Hoechst 33342 (blue) (C-a). 300 cells from 3 independent experiments were counted. Transfection efficiency of GFP-HDAC4 is 96%±3.64% (C-b). (D, E) The time-dependent changes in gene expression levels for aggrecan and type II collagen following CTS. Real-time PCR showed that mRNA level for aggrecan (D) and collagen II (E) were elevated after 2 and 3 h of CTS, but decreased after 4 h of CTS. Values are presented as mean±SD (n = 3). *P<0.05, **P<0.01 versus the non-stretched control group.