FIG 4.

Role of Beclin-1 in p17-induced lymphangiogenesis. (A) Immunoblotting analysis of Beclin-1 knockdown cells. Blots from one representative experiment out of three with similar results are shown (left). Values reported for Beclin-1 are the means ± SDs of three independent experiments (right). Statistical analysis was performed by one-way ANOVA; a Bonferroni posttest was used to compare data (***, P < 0.001). (B) LN-LECs were nucleofected with Beclin-1 and irrelevant (siScramble) siRNAs. Twenty-four hours after nucleofection, cells were serum starved for 16 h and then seeded on growth factor-reduced Cultrex BME in the presence or absence of 10 ng/ml of GST, p24, or p17. Pictures were taken after 12 h of culture on growth factor-reduced Cultrex BME (magnification, ×10). Values reported for tube formation are the means ± SDs of three independent experiments with similar results. Statistical analysis was performed by one-way ANOVA; a Bonferroni posttest was used to compare data (**, P < 0.01). (C) LN-LECs were conucleofected with EGFP-LC3 plasmid and siScramble or with EGFP-LC3 plasmid and siBeclin-1. Twenty-four hours after nucleofection, cells were incubated for 16 h in serum-starved medium and then left untreated or treated for 3 h with 10 ng/ml of GST, p24, or p17 in complete medium. The images display LC3 signals in green and cell nuclei in blue. Magnification, ×100. White arrows indicate dot-like LC3⁺ structure formation. NT, not treated.