Migration Assay

To evaluate the effect of blockade of IRS-1 on the migration of HCECs cells, a modified Boyden chamber assay was performed as described previously[33]. Briefly, 1×10⁴ cells in 100 µL DMEM medium were seeded in the upper chambers. After 24h incubation, the medium was replaced and fresh DMEM medium was added. Totally 500 µL of DMEM medium were added in lower chambers and treated with GS-101 for another 24h. The migrated HCECs were fixed prior to staining with 0.5% crystal violet solution. Non-invading cells were swapped with a cotton swab. The infiltrated cells were counted under a phase contrast microscopy. Each assessment of each experimental group was repeated several times.