Research Paper Volume 13, Issue 2 pp 2604—2625

Figure 5. Knockdown of CENPF inhibits proliferation, invasion and migration of LUAD cells via the ERβ2/5 pathway. (A) Immunofluorescence showed the co-localization of CENPF and ERβ in A549 and H1299 cells (400 x). (B) Cell proliferation assays of different grouped cells at specific times in A549 and H1299 cells. (C, D) Corresponding quantified histograms of migration and invasion in A549 and H1299 cells. The invasion and migration of cells in CENPF-KD+E2 group were significantly reduced when compared with CENPF-NC+E2 group. (E, F) Protein expression of MMP2, N-cadherin and E-cadherin in A549 and H1299 cells (The corresponding gray value are shown in Supplementary Figure 4B–4D): The expression of MMP2 and N-cadherin were significantly decreased in CENPF-KD+E2 group when compared with CENPF-NC+E2 group. (G) Scratch experiment showed that the migration of CENPF-KD+E2 group was significantly lower than CENPF-NC+E2 group in A549 and H1299 cells (P=0.000, 0.000). (H) Corresponding quantified histograms of the A549 cells at different stages of the cell cycle (G1, S and G2/M). (I) Protein expression of CCND1, CDK2 and CDK4 in A549 and H1299 cells (The corresponding gray value are shown in Supplementary Figure 4F). *P < 0.05. (J) Knockdown of CENPF inhibited the expression of ERβ2/5 in vitro. (The corresponding gray value are shown in Supplementary Figure 5A). (K) Protein expression of CENPF, ERβ, ERβ1, ERβ2 and ERβ5 in vitro experiment after treated with E2 and Ful treatment (The corresponding gray value are shown in Supplementary Figure 5C). *P < 0.05. P values were calculated with two-tailed unpaired Student’s t-test, or one-way analysis of variance.