Figure 3.

Knockdown of Capn4 reduces nasopharyngeal carcinoma (NPC)cell migration and invasion in vitro and in vivo. (a, b) 5-8F cells and CNE2 cells were stably transfected with constructs encoding siRNA specifically targeting Capn4 or scrambled sequence as siRNA control, after which semi-quantitative RT-PCR and Western blot analyses were performed to monitor mRNA and protein levels of Capn4 in Capn4 siRNA and control siRNA cells, as well as parental 5-8F cells and CNE2 cells. β-actin was probed as internal control. (c) Transwell assays were performed to measure in vitro migration and invasion of Capn4 siRNA and control siRNA cells. (d) To evaluate in vivo metastasis of Capn4 siRNA and control siRNA cells (n = 10 per group), 1 × 10⁶ cells were resuspended in 0.1 mL of PBS and injected via the lateral tail vein. After 10 weeks, mice were euthanized, and metastatic nodules in lung and liver were quantified using dissecting microscopy after H&E staining. Representative H&E staining images of lungs and livers were shown.