Additional file 1 of FK506 induces lung lymphatic endothelial cell senescence and downregulates LYVE-1 expression, with associated decreased hyaluronan uptake

Additional file 1: Supplementary Material and Methods (Matys et al., 2006). Supplementary Figure 1. Effect of 48H treatment with FK506 on LYVE-1. Western blot analysis of LYVE-1 and β-actin proteins in lung lymphatic endothelial cells treated without and with FK506 (10 ng/mL and 15 ng/mL) for 48H. Molecular weight (kDa) for each protein is indicated on the right. Ratio of LYVE-1 to β-actin density was expressed as fold-change relative to control. Data represent mean ± SEM of three independent experiments, consisting of one technical replicate each. Supplementary Figure 2. FK506 prevents NFAT nuclear translocation. Lung lymphatic endothelial cells were treated as indicated. Samples of protein lysates (Entry: 27%), and nuclear (27%) fractions were separated by SDS-PAGE and transferred to nitrocellulose membranes, and reacted with antibodies against NFAT, PARP (nuclear marker) and GAPDH (cytoplasmic marker). FK506 resulted in a marked decrease in NFAT in nuclear fraction. Data represent mean ± SEM of 3 independent experiments (p