Fig. 3

Sox9 enhances SHP transcription activity by binding to its promoter. A Heat map showed the differentially expressed genes in Sox9 f/f and H-Sox9 KO mice induced by IR. B Hepatic expression levels of Sox9 and SHP in Sox9 f/f and H-Sox9 KO mice were determined by qRT-PCR analysis following IR injury. 36B4 was used as a housekeeping gene. N = 4–5 per group, data are shown as mean ± SD. C SHP promoter contained a putative Sox9 binding site. Sequences of the WT (black) and mutant (red) binding sites. D Amplification and activity detection of different fragments of SHP promoter (position 40 to − 1500, 40 to − 1900 relative to the transcription start site). These fragments were inserted into the pGL3-Basic vector and transfected into Hepa 1–6 cells with or without Sox9 expression plasmids and pRL-TK using lipofectamine 2000. N = 3. E SHP-promoter or SHP-mutation promoter was co-transfected into Hepa1-6 cells with pRL-TK for 24 h and samples were analyzed by dual-luciferase assays. N = 3 per group. The ratio of firefly luciferase activity to renilla luciferase activity in control group was set to 1. F ChIP analysis showed that Sox9 interacted with classic binding site on SHP promoter. G ChIP-qPCR assay of Sox9 level on SHP promoter using chromatin solutions prepared from Sox9 f/f and H-Sox9 KO mouse livers. N = 3