After the introduction of pcDNA3.1-AFAP1-AS1 plasmids, AFAP1-AS1 shRNA or their NCs into Y-79 and WERI-Rb-1 cells, RT-qPCR analysis was performed to confirm the overexpression or silencing of AFAP1-AS1 in both the RB cells (Figure 2(c)). The gene discussed is RB1; the disease is retinoblastoma.