Fig. 4

lnc-DILC suppresses PTEN degradation. a The subcellular location of lnc-DILC in ccRCC cells. U6 and GAPDH were used as internal controls. b Biotinylated lnc-DILC or antisense lnc-DILC (AS-lnc-DILC) were incubated with ccRCC cell extracts, targeted with streptavidin beads, and associated proteins were resolved in a gel. Western blotting analysis of the specific association of PTEN and lnc-DILC. Antisense lncDILC was shown as a negative control. c RIP experiments were performed to examine the RNA associated with PTEN. HOTAIR and GAPDH mRNA was taken as negative control. d The PTEN mRNA levels in ACHN and 786-O cells transfected with pcDNC3.1-lnc-DILC or the empty vector. e The PTEN mRNA levels in Caki-1 cells transfected with lnc-DILC or negative control siRNAs. f The PTEN protein levels in ccRCC cells with lnc-DILC overexpression or knockdown were determined by western blot. g ACHN transfected with pcDNC3.1-lnc-DILC or the empty vector were treated with CHX (0.1 mg/ml) and harvested at the indicated times. Cells were lysed and cell lysates were then blotted with the indicated antibodies. Lower panel: quantification of the PTEN protein levels relative to GAPDH. h Caki-1 cells transfected with lnc-DILC or negative control siRNAs were treated with CHX (0.1 mg/ml) and harvested at the indicated times. Cells were lysed and cell lysates were then blotted with the indicated antibodies. Lower panel: quantification of the PTEN protein levels relative to GAPDH. *p < 0.05, **p < 0.01, ***p < 0.001