Fig 2. GSK3 inhibition induces proliferation, migration and network formation of primary human lung lymphatic endothelial cells.

A) Primary human lung LECs in 1% FBS were incubated with SB216763 (1μM) or vehicle (DMSO) alone. Cell proliferation was determined at 48h after treatment using CyQUANT assay (A). Effect of SB216763 (1μM) on the migration of primary LECs was evaluated in transwell migration assays (B). Effect of SB216763 (1μM) on tube formation of primary LECs. An equal number of LECs were seeded onto growth factor-reduced Matrigel at a density of 1.2x10⁴ cells/well and incubated with either SB216763 (1μM) or vehicle alone (DMSO). After 16 hours, cells were labeled with Calcein-AM and random 4X fluorescent images were taken (C) and the number of meshes (D) and mesh area (E) per image were determined using ImageJ (NIH) Angiogenesis Analyzer plugin. Results are expressed as fold change compared to vehicle control. Data represent Mean ± SEM of a single experiment. All experiments were repeated a minimum of three times. *P < 0.05, ** P < 0.01, *** P < 0.001 by t-test.