Fig. 6

Validation of protein–protein interaction network by siRNA knockdown and CCK-8 assays. a Gene expression levels of TGFB1, RHOA, ILK, CDC42, LYN and FLNA validated by qRT-PCR under TGFB1 silencing in primary chondrocytes. Relative mRNA levels were normalized to the GAPDH level and calculated using the 2^−ΔΔCT method. The relative fold-changes of each gene in the TGFB1 siRNA group were normalized and compared to the Negative siRNA group, separately. Data are presented as the mean with standard deviation for technical triplicates in an experiment. * represents p < 0.01, and *** represents p < 0.001 in a t-test for the difference in gene expression level. b Effects of GZZSZTW-medicated serum on the proliferation of primary chondrocytes under TGFB1 silencing. CCK-8 assay was used to detect the chondrocyte proliferation following treatment with GZZSZTW-medicated serum at increasing concentrations (0%, 5%, 10% and 20%) for 24 h after siRNA knockdown. The ordinate represents the cell viability (%), while the abscissa represents the concentration of GZZSZTW-medicated serum (%) in the culture of primary chondrocytes. Cell viability was normalized and calculated relative to that of the blank group (0%). Data are presented as the mean of three independent experiments for technical triplicates with standard deviation. Δ and ΔΔ represent p < 0.05 and p < 0.01, and ### represents p < 0.001 in a t-test for the difference in cell viability (%)