Fig 2: HOXC8 knock-down inhibited the chondrogenic differentiation of SCAPs. a, b Alcian Blue staining and quantitative analysis results showed that knock-down of HOXC8 reduced chondrogenic differentiation in SCAPs. c–e Real-time RT-PCR results showed that knock-down of HOXC8 downregulated the expressions of COL2 (c), COL5 (d), and SOX9 (e) in SCAPs. f Alcian Blue and Picro Sirius Red staining results of chondrogenesis induced pellet. Scale bar, 100 µm. GAPDH was used as an internal control. Student’s t test was performed to determine the statistical significance. All error bars represent s.d. (n = 3). *P = 0.05. **P = 0.01. Consh, control shRNA; HOXC8sh, HOXC8 shRNA.

Fig 3: DLX5 over-expression enhanced the chondrogenic differentiation in SCAPs. a DLX5 expression during chondrogenic differentiation as detected by real-time RT-PCR. b, c Real-time RT-PCR and Western blot results showed the DLX5 expression in SCAPs. The relative protein levels quantified by densitometry and normalized to GAPDH. d, e Alcian Blue staining and quantitative analysis results show that DLX5 over-expression enhanced chondrogenic differentiation in SCAPs. f–h Real-time RT-PCR results show that DLX5 over-expression upregulated the expression of COL2 (f), COL5 (g), and SOX9 (h) in SCAPs. i Alcian Blue and Picro Sirius Red staining results of chondrogenesis induced pellet. Scale bar, 100 µm. GAPDH was used as an internal control. One-way ANOVA or Student’s t test analysis was performed to determine the statistical significance. All error bars represent s.d. (n = 3). *P = 0.05. **P = 0.01

Fig 4: HOXC8 over-expression enhanced the chondrogenic differentiation of SCAPs. a, b Alcian Blue staining and quantitative analysis results showed that HOXC8 over-expression enhanced chondrogenic differentiation in SCAPs. c–e Real-time RT-PCR results showed that HOXC8 over-expression upregulated the expression of COL2 (c), COL5 (d), and SOX9 (e) in SCAPs. f Alcian Blue and Picro Sirius Red staining results of chondrogenesis induced pellet. Scale bar, 100 µm. GAPDH was used as an internal control. Student’s t test was performed to determine the statistical significance. All error bars represent s.d. (n = 3). **P = 0.01

Fig 5: DLX5 knock-down inhibited the chondrogenic differentiation in SCAPs. a, b Real-time RT-PCR and Western blot results showed that DLX5 was knocked down in SCAPs. The relative protein levels quantified by densitometry and normalized to GAPDH. c, d Alcian Blue staining and quantitative analysis results show that knock-down of DLX5 reduced chondrogenic differentiation in SCAPs. e–g Real-time RT-PCR results show that knock-down of DLX5 downregulated the expressions of COL2 (e), COL5 (f), and SOX9 (g) in SCAPs. h Alcian Blue and Picro Sirius Red staining results of chondrogenesis induced pellet. Scale bar, 100 µm. GAPDH was used as an internal control. Student’s t test was performed to determine the statistical significance. All error bars represent s.d. (n = 3). **P = 0.01. Consh, control shRNA; DLX5sh, DLX5 shRNA.