Fig. 7

Gelsolin shRNA decreases cell invasion and migration in canine OSA8 cells. a Protein lysates were generated from canine OSA8 cells stably transduced with either pLK0.1-scramble-hygroB (Scr) control vector or pLK0.1-shGSN-hygroB (shGSN) lentivirus and positive clones were selected for with Hygromycin B. Protein lysates were separated via SDS-PAGE and western blotting for gelsolin (GSN) and β-actin was performed to confirm efficiency of GSN knockdown. b Canine OSA8 cells expressing pLK0.1-shGSN-hygroB or scramble control were collected and real-time PCR for gelsolin was performed (Bars: SD. Statistical analysis: one-way ANOVA, *p < 0.01). c The invasive capacity of OSA8 cells transduced with either pLK0.1-scramble-hygroB control vector or pLK0.1-shGSN-hygroB lentivirus was evaluated using standard Matrigel invasion assays. Cells (5 × 10⁴) were plated in serum free medium and transferred onto cell culture inserts coated with Matrigel® for 24 h. After incubation, cells remaining on the upper surface of the insert membrane were wiped away using a cotton swab, and cells that had migrated to the lower surface were stained with crystal violet and counted in ten independent 20x hpf for each sample. Three independent experiments were performed using cells from 3 separate transduction experiments and all reactions were performed in triplicate