Islet culture improves islet viability by increasing ATP content and OCR.

<p><i>(</i><b><i>a</i></b><i>)</i> ATP was measured and normalized to DNA content (nmol/mg protein/mg-DNA) as described in <i>“</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099796#s2" target="_blank"><i>Materials and Methods</i></a><i>”</i>. Graphic represents ATP/DNA values. Results are means ± SEM of five separate experiments performed in triplicate (n = 5, <i>^**P<0.01</i> for <i>D₀</i> (black bars) <i>vs. D₇</i> (grey bars)). <i>(</i><b><i>b</i></b><i>)</i> Islet samples were used for OCR measurements normalized to DNA content (nmolO₂/min/mg-DNA) as described in <i>“</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099796#s2" target="_blank"><i>Materials and Methods</i></a><i>”</i>. Graphic represents OCR/DNA values. Data are means ± SEM of eighteen separate experiments performed in triplicate (n = 18, <i>^***P<0.001</i> for <i>D₀ vs. D₇</i>). <i>(</i><b><i>c/d</i></b><i>)</i> Protein extracts (30 µg) from islet samples with <i>(</i><b><i>c</i></b><i>)</i> high or <i>(</i><b><i>d</i></b><i>)</i> low JNK3 protein content in <i>D₀</i> and <i>D₇</i> were tested by Western blot. Polyclonal antibodies against: p-Akt1, p-Akt2, p-GSK3β, JNK3, MKK7, Akts and PTEN were used. Equal protein loading was assessed by blotting membranes with antibodies against tubulin and GAPDH. Data are representative of at least three separate experiments (n = 3–8).