Fig. 3

LncRNA uc.134 binds CUL4A and inhibits CUL4A nuclear export. a LncRNA uc.134-associated proteins identified by RNA pulldown and mass spectrometric analyses in HEK293T cells (upper panel). Western blot analysis of pulldown product (bottom panel). b RIP assays in HEK293T cells. qRT-PCR analysis of RIP (upper panel). Agarose electrophoresis of PCR products (bottom panel). Experiments were performed in triplicate, and data are presented as mean ± SD. ***P < 0.001. c Immunoblot (IB) detection of CUL4A retrieved by in vitro transcribed biotinylated RNAs corresponding to different fragments of uc.134 or its antisense sequence (dotted line) in HEK293T cells. d IB detection of Myc-tagged CUL4A (WT versus domain truncation mutants) precipitated by in vitro transcribed biotinylated uc.134 in HEK293T cells. Upper panel: graphic illustration of the domain structure of CUL4A. e–f LncRNA uc.134 inhibits CUL4A nuclear export as demonstrated by immunofluorescence staining (e) and Western blotting (f)