Fig. 4: Effects of LRP1 knockdown on the migration, invasion, and signaling of esophageal squamous cell carcinoma (ESCC) cells.

a, b ESCC cells were transfected with 20 nM siRNA targeting LRP1 (siLRP1). Negative control siRNA (siNC) was transfected into ESCC cells as negative control. LRP1 knockdown was confirmed by (a) RT-PCR with GAPDH as the internal control, and by (b) quantitative RT-PCR with LRP1 normalized to levels in respective samples and ACTB (β-actin) as the internal control. c Representative western blots of LRP1 knockdown in ESCC cells. β-actin used as the loading control. d Transwell assay of the effects of LRP1 knockdown on the migration of TE-8, TE-9, and TE-15 cells with or without 10 ng/mL rhPAI-1. Migrating cells were counted in four randomly chosen fields. e Transwell assay of the effects of LRP1 knockdown on the invasion of TE-8, TE-9, and TE-15 cells with or without 10 ng/mL rhPAI-1. Invading cells were counted in four randomly chosen fields. f Representative western blots of the total protein of LRP1-silenced ESCC cells using specific antibodies against Akt, p-Akt (Ser473), p-Akt (Thr308), Erk1/2, p-Erk1/2 (Thr202/Tyr204), and β-actin compared to the control. For (b), (d), and (e), data represent the mean ± SEM of triplicate wells for three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001).