Since the profiling of the 5hmC level was indirect, which was calculated by subtracting oxRRBS methylation from RRBS methylation levels, we performed 5hmC-specific PCR to validate one of the most hypermethylated 5hmC DMRs that is annotated to TUBG1, and a top hypomethylated 5hmC DMR annotated to EZHIP. Consistent with the sequencing results, we observed that the 5hmC level of TUGB1 was significantly high in recurrent bladder samples, while the 5hmC level of EZHIP was decreased. This evidence concerns the gene TUBG1 and carcinoma.