Fig. 4

LncRNA uc.134 inhibits the CUL4A-mediated ubiquitination of LATS1 to regulate Hippo kinase signaling. a–b Hierarchical clustering analysis (a) and GO analysis (b) of microarray data for HCC cells overexpressing uc.134 and control cells. Upregulated genes are highlighted in yellow, and downregulated genes are in blue. (fold changes >3; P < 0.05). The top ten most significantly enriched GO terms in differentially expressed genes relative to all other genes in the genome are shown. c IP experiment. d The overexpression of uc.134 prolonged the half-life of LATS1. Immunoblot detection of LATS1 (left); IB data were quantified using the ImageJ software (right). Error bars indicate the mean ± SD. **P < 0.01. e Ubiquitination assays of cells transfected with uc.134 plasmid in Bel7402 cells (left) or siRNA in QGY7701 cells (right). The bottom panels depict the input of the cell lysates. f Western blotting in Bel7402 cells showed that uc.134 reversed the CUL4A-mediated inhibition of Hippo kinase activity. g Western blots showed that the overexpression of uc.134 activates Hippo kinase signaling (left), whereas silencing of uc.134 yielded opposite results (right). h qRT-PCR analysis of cells overexpressing uc.134 or co-transfected with YAP plasmids compared with vector controls. Experiments were performed in triplicate, and data are presented as the mean ± SD. *P < 0.05; **P < 0.01; and ***P < 0.001