Figure 6.

Stimulation of S100A8 and S100A9 protein expression in naïve primary keratinocytes. Primary keratinocytes were isolated from wildtype C57BL/6 mice, matured, and stimulated for 24 h ex vivo. (A) Messenger RNA (mRNA) expression of S100A8 and S100A9 were analyzed by quantitative real-time PCR. Columns represent means ± SEM of at least five independent experiments. Values were standardized to RPL housekeeping gene and represent the n-fold to their respective unstimulated WT keratinocytes at each maturation time point (w/o = 1). Asterisks indicate significant differences to unstimulated controls based on the original expression data (ANOVA *p < 0.05; **p < 0.01; ***p < 0.001). (B) Release of S100A8/S100A9 into culture supernatants by WT keratinocytes during maturation was analyzed after incubation with different stimuli via ELISA. Columns represent the means ± SEM of at least three independent experiments. Values represent the n-fold to unstimulated WT keratinocytes (w/o = 1). Student’s t-test revealed no significant differences. (C) Expression levels of S100A8 and S100A9 in primary WT keratinocytes were determined by Western blotting. β-actin served as loading control. One representative experiment out of two independent experiments is shown.