Figure 3.

E2F1 augments NSCLC cell viability, migration, and invasion through transcriptionally activating NELL2

(A) Boxplot of NELL2 expression in lung adenocarcinoma samples from TCGA database. (B) Correlation between the expression of E2F1 and NELL2 in lung adenocarcinoma samples. (C) qRT-PCR to determine NELL2 expression in NSCLC cell lines (A549, NCI-H1299, NCI-H23, NCI-H460) and human lung epithelial cell line 16HBE. (D) ChIP assay to verify the interaction between the E2F1 and NELL2 promoter. (E) Dual-luciferase reporter gene assay to verify the binding between NELL2 and E2F1. (F) qRT-PCR to determine NELL2 expression in the presence of E2F1 overexpression/silencing in A549 cells. (G) Western blot to determine NELL2 expression in the presence of E2F1 overexpression/silencing in A549 cells. (H) qRT-PCR to determine the expression of E2F1 and NELL2 in A549 cells of each group. (I) Western blot to determine the expression of E2F1 and NELL2 in A549 cells of each group. (J) CCK-8 assay to detect the viability of A549 cells of each group. (K) EdU staining to detect the viability of A549 cells of each group. (L) Wound-healing assay to detect the migration of A549 cells of each group. (M) Transwell to assess the invasion of A549 cells of each group. ∗p < 0.05. One-way ANOVA was applied for comparison among multiple datasets and two-way ANOVA for comparison among data of multiple datasets at different time points. Each experiment was conducted in triplicates.