Fig. 1

LATS1 is downregulated and interacts with circXRN2 in bladder cancer. a LATS1 was downregulated in surgical tissue samples (n = 30). Scale bar: 200 μm. b RIP assay followed by RNA sequencing revealed that LATS1 protein could interact with numerous circRNAs. c CircRNAs binding with LATS1 were validated by qRT‒PCR in different cell lines (SV-HUC-1, 5637, EJ, T24, TCCSUP and UM-UC-3). d The affinities of dysregulated circRNAs to the LATS1 protein were evaluated by RIP assay, and the results showed that circXRN2 had the highest affinity for LATS1. e Total RNA was extracted from clinical specimens of our own cohort (n = 40). Dysregulation of circXRN2 in surgical tissue samples was confirmed by qRT‒PCR. f The expression levels of LATS1, p-TAZ, TAZ, p-YAP and YAP in cell lines were determined by western blotting. g Immunofluorescence indicated the localization of TAZ/YAP in cell lines with various abundances of circXRN2. Scale bar: 50 μm. h The biological structure of circXRN2 is represented by a graphic illustration, and the back-splicing site was verified by PCR and Sanger sequencing. i CircXRN2 was only amplified by divergent primers in cDNA but not in gDNA of T24 and TCCSUP cells. GAPDH was used as a negative control. j Cells were incubated in the absence or presence of RNase R, and the expression levels of circXRN2 and linear XRN2 were determined by qRT‒PCR. k RNA FISH indicated that CircXRN2 was mainly located in the cytoplasm. The nuclei were stained with DAPI, while circXRN2 was red. Scale bar: 50 μm. All the data are presented as the mean ± standard deviation (n = 3). *P < 0.05, **P < 0.01, compared with the control group