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. 2015 May 13;56(5):2985–2992. doi: 10.1167/iovs.15-16466

Figure 4.

Figure 4

Glucolipotoxicity increases Nox2 and Rac1, and these two proteins are colocalized. Localization of Nox2 and Rac1 was performed immunohistochemically in cells incubated in 5 mM or 20 mM glucose for 48 hours, in the presence or absence of palmitate or ceramide, supplemented with NSC23766. For Nox2 (green) FITC-conjugated, and for Rac1, Texas Red–conjugated secondary antibodies were used, and the coverslips were mounted using Vectashield with DAPI (blue): 5 mM Glu, 5 mM Glu+Palm, and 5 mM Glu+Cer = cells incubated in 5 mM glucose with no addition or with 50 μM palmitate or ceramide, respectively; 20 mM Glu, 5 mM Glu+Palm, and 20 mM Glu+Palm+NSC = cells incubated in 20 mM glucose with no addition or with palmitate supplemented with or without 20 μM NSC23766; 20 mM Glu+Cer and 20 mM Glu+Cer+NSC = cells incubated in 20 mM glucose and 50 μM ceramide in the absence or presence of NSC23766, respectively.