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Figure 7


Fig. 7. Insulin-induced STAT3 nuclear translocation is abrogated by PKC{delta} inhibition. Primary keratinocytes were plated on glass slides and maintained for 5 days in low-Ca2+ MEM (0.05 mM) until they reached 80% confluence. (A) Cells were left untreated (upper panel) or pre-treated with 5 µM rottlerin (Rott) for 7 minutes (lower panel), followed by 10–7 M insulin for 5 and 30 minutes (Ins). Cells were fixed in methanol, washed and air-dried. Cultures were analyzed by immunofluorescence using anti-phospho-tyr-705-STAT3 (STAT3-p-tyr) antibody, followed by FITC-conjugated secondary antibody. Slides were viewed by confocal microscopy. (B) Cells were untreated (–) or treated with insulin (Ins) for 5 and 30 minutes. Following treatment, cells were fixed in methanol and analyzed by immunofluorescence using anti-PKC{delta} antibody followed by FITC conjugated secondary antibody and scanned by confocal microscope. Arrow in middle panel indicates translocation of PKC{delta} to the perinuclear membranes. Experiments were repeated at least three times. Bar, 20 µm.





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