Long noncoding RNA semaphorin 6A-antisense RNA 1 reduces hepatocellular carcinoma by promoting semaphorin 6A mRNA degradation

Figure 4 Semaphorin 6A-antisense RNA 1 combined with semaphorin 6A mRNA downregulates semaphorin 6A expression. A and B: The expression levels of semaphorin 6A (SEMA6A) in Hep3B and LM3 cells stably transfected with pc-SEMA6A-AS1 or pcDNA3.1 plasmid were assessed by quantitative polymerase chain reaction (qPCR) for mRNA (A) and Western blot for protein (B); C: Transcript location of SEMA6A-AS1 and SEMA6A; D: Schematic of the tagged RNA affinity purification experiment; E: The level of SEMA6A mRNA in RNAs obtained by tagged RNA affinity purification experiment was analyzed by qPCR; F: The direct binding of SEMA6A-AS1 and SEMA6A mRNA in Hep3B cells was assayed by luciferase activity with luciferase reporter vector containing the binding site or mutant-form binding site; G: The levels of remaining SEMA6A mRNA at each timepoints after actinomycin D treatment were detected by qPCR. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. SEMA6A: Semaphorin 6A; TRAP: Tagged RNA affinity purification; Mut-BS: Mutant-form binding site; GST: Glutathione-S-transferase; RBPs: RNA binding proteins; qPCR: Quantitative polymerase chain reaction.