Interleukin 6 Function in the Skin and Isolated Keratinocytes Is Modulated by Hyperglycemia

Figure 4

High glucose alters STAT3 activation in cultured keratinocytes. Neonatal mouse keratinocytes (MEKn) were cultured for 7 days in either normal glucose (NG; 5 mM), high glucose (HG; 25 mM), or mannitol osmotic control (Man; 5 mM mannitol). (a) Basal IL-6Rα mRNA expression was determined via qPCR. Data are presented as the ; (3 experiments). (b) MEKn were incubated with 0 or 5 ng/ml of rmIL-6 for 30 minutes. STAT3 phosphorylation (Y705) was determined via Western blotting (left panel) and quantified via densitometry analysis (right panel). (c) MEKn were treated with 0, 5, or 20 ng/ml of rmIL-6 for 30 minutes and stained for pSTAT3 (Y705) (green) and nucleus (blue). (d) MEKn were stained for SOCS3. Percentage of positive staining for pSTAT3 or SOCS3 was determined Operetta High-Content Imaging System (PerkinElmer, Waltham, MA) and analyzed with Columbus (v2.6.0, PerkinElmer). Data are presented as the ; . and , significantly different from normal glucose-treated cells.

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