Fig. 6: Effects of ManN on non-ECs.

Effects of ManN on non-endothelial cells of bovine, mouse, or human origin. ManN did not promote growth of Calu6 (a), A673 (b), U87MG (c), and 4T1 (d) tumor cells. Ten-percent FBS was used as positive control for Calu6 and A673, whereas 10 ng/ml bFGF and 1 μg/ml human apo-transferrin were used as positive controls for U87MG and 4T1, respectively. Similarly, no increases in proliferation were induced by ManN on AML12 (e), bovine pituitary cells (f), NIH3T3 cells (g), human RPEs (h), human dermal fibroblasts (i), and human keratinocytes (j), alone or in combination with growth factors. Proliferation quantification was performed using AlamarBlue® or MTS (for 4T1 cells). n = 3 independent samples. Inserted are representative western blot analyses showing dose-dependent effects of ManN and mannose at 400 μM (2,4) and 2 mM (3,5) on bFGFR1 or β1 integrin (for 4T1, AML12, NIH3T3 cells, human skeletal muscle cells, human dermal fibroblasts, and human keratinocytes) molecular mass compared to the untreated control (1). β-actin served as loading control. GM: growth media. Proteins were separated on NuPAGE 3–8% Tris-Acetate gel for western blot analysis. For each study, a representative experiment is shown from two independent studies. Asterisks indicate a significant difference compared with control. When statistical analysis was done using a different control, bracket was used between specified groups. Data were means +/− SD of the mean or an average when n = 2. Statistical analysis was done by two-tailed, two-sample unequal variance t test. *p < 0.05, **p < 0.01. Data are provided as a Source data file.