Research Paper Volume 12, Issue 20 pp 20396—20412

Figure 3. FAM83C-AS1 inhibits SEMA3F expression through interacting with EZH2 in CRC tumor cells. (A) The interaction between FAM83C-AS1 and SEMA3F promoter was tested by luciferase reporter assay. (B) SEMA3F was speculated to boast the potential of binding with EZH2 via UCSC. (C, D) H3K27me3 occupancy and EZH2 binding of the SEMA3F promoter in transfected cells were analyzed using ChIP. (E) The efficiency of EZH2 knockdown was detected using RT-qPCR. (F) The interaction between EZH2 and SEMA3F promoter was investigated using luciferase reporter assay. (G) RT-qPCR and western blotting assays were conducted to detect the mRNA and protein expression of SEMA3F after RKO and SW620 cells were transfected with sh-EZH2#1 or sh-NC. (H) RIP assay demonstrated that SEMA3F could bind to EZH2 in RKO and SW620 cells. (I) RT-qPCR and western blotting analyses were used to detect the mRNA and protein expression of EZH2 in RKO and SW620 cells transfected with sh-FAM83C-AS1#1 or sh-NC. (J) FAM83C-AS1 expression in RKO and SW620 cells transfected with sh-EZH2#1 or sh-NC was determined using RT-qPCR. (K) H3K27me3 occupancy and EZH2 binding to SEMA3F promoter in transfected cells were analyzed using ChIP. (L) mRNA and protein expression of SEMA3F in RKO and SW620 cells transfected with different plasmids were examined using RT-qPCR and western blotting analysis. ^**P < 0.01. n.s.: no significance.