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. 2011 Oct 17;60(11):2830–2842. doi: 10.2337/db11-0347

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© 2011 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 1. — Effect of high glucose on AA and LA content in INS-1E cells. A: INS-1E cells were incubated with the indicated glucose (Glc) levels for 16 h and processed for analysis of fatty acid residues in membrane phospholipids. The absolute content of AA and LA in phospholipids of INS-1E cells exposed to 5, 11, and 25 mmol/L glucose was calculated as follows: , where A_STD is the area of the standard reference of the fatty acid from the calibration run, C_STD is the concentration of the standard reference from the calibration run, A_x is the area of the compound (x) from the calibration run, C_x is the concentration of the compound (x) from the calibration run, F_x is the conversion factor of the compound (x), C_GC is the concentration of the compound in the sample from the GC trial run, A_GC is the area of the compound (x) obtained from the GC trial run, and MW_x is the molecular weight of the compound (x). Results are mean ± SEM, n = 4. *P < 0.05 for the difference from the respective 5 mmol/L glucose values. B: Lysates were prepared from similarly treated INS-1E cells and taken for Western blot analysis of total cPLA₂, pSer⁵⁰⁵-cPLA₂ (white bars), and pSer⁵¹⁵-cPLA₂ (black bars). Representative Western blots are shown (inset). Tubulin was used for equal protein loading control. Results are mean ± SEM, n = 3. *P < 0.05 for the difference from the respective 5 mmol/L glucose control.

Inline graphic — Effect of high glucose on AA and LA content in INS-1E cells. A: INS-1E cells were incubated with the indicated glucose (Glc) levels for 16 h and processed for analysis of fatty acid residues in membrane phospholipids. The absolute content of AA and LA in phospholipids of INS-1E cells exposed to 5, 11, and 25 mmol/L glucose was calculated as follows: , where A_STD is the area of the standard reference of the fatty acid from the calibration run, C_STD is the concentration of the standard reference from the calibration run, A_x is the area of the compound (x) from the calibration run, C_x is the concentration of the compound (x) from the calibration run, F_x is the conversion factor of the compound (x), C_GC is the concentration of the compound in the sample from the GC trial run, A_GC is the area of the compound (x) obtained from the GC trial run, and MW_x is the molecular weight of the compound (x). Results are mean ± SEM, n = 4. *P < 0.05 for the difference from the respective 5 mmol/L glucose values. B: Lysates were prepared from similarly treated INS-1E cells and taken for Western blot analysis of total cPLA₂, pSer⁵⁰⁵-cPLA₂ (white bars), and pSer⁵¹⁵-cPLA₂ (black bars). Representative Western blots are shown (inset). Tubulin was used for equal protein loading control. Results are mean ± SEM, n = 3. *P < 0.05 for the difference from the respective 5 mmol/L glucose control.