Fig. 6: COL18A1-AS1/KLF12 axis repressed ccRCC progression through UCP1-mediated lipid browning.

A, B qRT-PCR and Western blotting assays showed the expression level of KLF12 in CAKI-1 and OS-RC-2 cells co-transfected with COL18A1-AS1 vector and/or si-KLF12. C, D Cell proliferation ability of OS-RC-2 cells co-transfected with COL18A1-AS1 vector and/or si-KLF12 was determined using CCK8 assays or colony formation assays. E, F Cell migration and invasion ability of OS-RC-2 cells was measured with Transwell assays or wound healing assays (Magnification: ×100 for Transwell assays and x40 for wound healing assays). G Photomicrographs of Oil Red staining of OS-RC-2 cells. Scale bars, 10 μm. H Relative TG (mmol/gprot) levels in OS-RC-2 cells assessed by a triglyceride assay kit. I Representative photographs of IF staining for lipids with BODIPY 493 of 503 (green) and KLF12 (red) in the cells co-transfected with COL18A1-AS1 vector and/or si-KLF12. Cell nuclear appear in blue (DAPI). Scale bars, 10 μm. J Protein levels of lipid browning marker genes (UCP1, PGC1A, CIDEA, and DIO2) in CAKI-1 and OS-RC-2 cells co-transfected with COL18A1-AS1 vector and/or si-KLF12 assessed by western blotting. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars indicate mean ± SD. All the experiments were performed in triplicate.