Figure 6.

qRT‐PCR analyses of gene transcription. (A) Unchanged expression of genes coding for key regulators involved in Wnt/β‐catenin signalling pathway (Axin‐1, β‐cat, Dvl‐1, Gsk‐3β and Hif‐1α) and upregulated expression of genes coding for transcription factor SOX9 (Sox9), the cytokine CYTL‐1(Cytl‐1) and genes coding for chondrocyte proteins of collagen type II (Col2‐α1) and Aggrecan (Acan) in 28.0 μM Sal B‐treated chondrocytes. (B) qRT‐PCR analyses demonstrate actions of various concentrations of Sal B on expression of Sox9, Cytl‐1, Col2‐α1 and Acan in cultured chondrocytes in a similar dosage effective fashion, by which the expression of these genes was significantly upregulated by a series of dilutions of Sal B accept the high concentrations of 28.0 and 42.0 μM for Cytl‐1. (C) The expression of Col1‐α1, Col2‐α1 and Acan in chondrocytes treated with 28.0 μM Sal B in a time course of 6, 12, 24 and 48 hrs was investigated by qRT‐PCR. The expression of Col2‐α1 and Acan was remained upregulated in the testing period accept 12 hrs for Acan, while Col1‐α1 was not stimulated by Sal B in the analysis even using six times cDNA templates beside an increase at 24 hrs. (D) The time course study for the expression of cytokine‐like factor Cytl‐1 and interleukin‐1β (IL‐1β). The IL‐1β was highly upregulated by Sal B treatment at 6 hrs and then became unchanged, while the Cytl‐1 expression remained unchanged in the early hours and upregulated at 24 hrs and 48 hrs. The house‐keeping gene GAPDH was used as an internal reference. *P < 0.05 and **P < 0.01.