Fig. 3

TMEM147 upregulates 27HC by activating DHCR7, and promotes the progression of HCC.(a) RNA-seq revealed the differentially regulated genes between TMEM147-silenced cells and control cells. (b) Gene ontology (GO) analysis revealed that the differentially expressed genes were significantly enriched in cholesterol metabolism pathways. (c) Protein levels of key enzymes in cholesterol metabolism pathways in TMEM147-silenced and control Huh7 cells. (d) DHCR7 protein level of TMEM147 overexpression, silenced, and control cells. (e) DHCR7 mRNA level of TMEM147 overexpression, silenced, and control cells. (f) Cholesteryl esters (CEs) level of TMEM147 overexpression, silenced, and control HCC cells, and different TMEM147 expression subcutaneous tumors. Analyzed by ELISA assay. (g) 27HC level of TMEM147 overexpression, silenced, and control HCC cells, and corresponding cell culture medium. Analyzed by ELISA assay. (h) CYP27A1 protein level of TMEM147 overexpression, silenced, and control cells. (i) Growth curve assay based on CCK8 analysis in TMEM147 overexpression HCC cells with different condition, including DHCR7 silencing, CYP27A1 inhibiting, and addition of exogenous 27HC. (j) Representative images of Transwell migration and Matrigel invasion assay for the indicated cells. Scale bars, 100 μm