Research Paper Volume 12, Issue 21 pp 21674—21686

Circular RNA circSEMA5A promotes bladder cancer progression by upregulating ENO1 and SEMA5A expression

Figure 7. CircSEMA5A accelerates BC angiogenesis through upregulating SEMA5A. (A and B) qRT-PCR and western blot assays detecting SEMA5A expressions after T24 cells were transfected with circSEMA5A overexpression plasmids and UM-UC-3 cells were transfected with circSEMA5A siRNAs. (C and D) Angiogenesis capability was assessed by tube formation assay after T24 cells were transfected with circSEMA5A overexpression plasmids together with SEMA5A siRNAs and UM-UC-3 cells were transfected with circSEMA5A siRNAs together with SEMA5A overexpression plasmids. (E) Binding motif of EIF4A3 in circSEMA5A and SEMA5A mRNA. (F and G) RIP validated the interactions of EIF4A3 with circSEMA5A and SEMA5A mRNA. (H and I) Cells were transfected with circSEMA5A or EIF4A3 overexpression plasmids or siRNAs, the enrichments of SEMA5A mRNA in anti-EIF4A3 precipitates were analyzed by qRT-PCR and compared with NC group. (J and K) The influence of circSEMA5A or EIF4A3 on the stability of SEMA5A mRNA was assessed by RNA stability analysis. Data are presented as mean ± SD. NS, no significance; *P < 0.05.