FIGURE 5.

AKT and ERK phosphorylation in ASCs under normoxia and hypoxia in patient group B. Group B included adipose tissue samples from chronic non-healing wounds, which exhibited high MIF secretion in the hypoxic culture of ASCs. Cells were cultured in growth medium at normoxic oxygen levels of 21% O₂ or hypoxic oxygen levels of <1% O₂. Cell lysates were obtained at indicated measuring points and Western blot analysis was performed and evaluated using the AIDA software. Tubulin was detected for total protein standardization. (A) Blots are representative of all Western blots performed to detect levels of phosphorylated AKT (pAKT), AKT, phosphorylated ERK (pERK), and ERK. Analyses were performed for three biological replicates (n = 3). The graphs display (B) relative pAKT/AKT and (C) relative pERK/ERK were normalized to the baseline value at 0 h, set to one. Data are mean values ± SEM (two-way ANOVA, Holm-Šídák’s multiple comparisons test, ^∗p < 0.05).