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. 2013 Jan;27(1):199–207. doi: 10.1096/fj.12-211896

Figure 1.

Figure 1.

Chiro-inositols stimulate upstream effectors of insulin signaling in hippocampal neurons. A) Western blot analysis of IRβ pTyr in hippocampal neurons after treatment with insulin (1 μM) or chiro-inositols (DCI, pinitol, or INS-2; 100 μM) shows an increase of ∼2.5 fold with insulin (2.58±0.06) compared to control. Similar results were observed after treatment with DCI (2.28±0.18), pinitol (2.47±0.10, shown), and INS-2 (2.40±0.42). B) Akt phosphorylation (Thr308) in hippocampal neurons treated with insulin (1 μM) or inositols (DCI, pinitol, or INS-2; 100 μM). Using immunofluorescence microscopy, we observed a primarily somatic distribution of pAkt. Treatment with insulin increased pAkt levels ∼1.7-fold (1.69±0.16). Treatment with chiro-inositols (DCI shown) produced a ∼1.5-fold (1.53±0.21) increase in pAkt levels. Similar increases in pAkt levels relative to loading control (cyclophilin B) were observed by Western blot, with both insulin and inositols producing ∼1.3-fold increases in Akt phosphorylation. C) Immunocytochemical detection of pIRS-1 (Ser307) in hippocampal neurons treated with insulin (1 μM) or chiro-inositols (DCI, pinitol, or INS-2; 100 μM). Insulin treatment increased IRS-1 phosphorylation by ∼2-fold (1.87±17). Chiro-inositols (DCI shown) also increased IRS-1 phosphorylation ∼2-fold (2.02±19). IB, immunoblot; IP, immunoprecipitation; a.u., arbritrary units. *P < 0.05, **P < 0.01, ***P < 0.001; unpaired t test.