Figure 3. Knockdown of TPT1-AS1 inhibits the VEGFA expression by decreasing VEGFA mRNA stability. (Ai) Flexmap liquichip assays were performed to detect the metastasis-associated factors in control or sh-TPT1-AS1 HCT116 cell supernatant. The results were also shown in heatmap (Aii). (B) TCGA date was collected to analyze the relationship between TPT1-AS1 and VEGFA expression. (C) Western blot assays and qRT-PCR were used to test the VEGFA expression in indicated CRC cells. (D) Combined IF/FISH assays showed that TPT1-AS1 was mainly expressed in cytoplasm of CRC cells and TPT1-AS1 expression positively correlated to VEGFA expression. (E) Luciferase reporter assays were used to detect the possibility that TPT1-AS1 transcriptionally regulated VEGFA expression. (F) VEGFA mRNA levels were detected using actinomycin D mRNA stability assays in indicated CRC cells. *P < 0.05, **P < 0.01, # represents no statistical significance.