Group data–between-subject variation (mean concentrations, 95% confidence limits) of intact proinsulin (upper panel), insulin (middle panel) and the proinsulin:insulin ratio (lower panel) at collection (0h) and at the five time points thereafter, in samples kept at 4°C (left-hand panels) and at room temperature (right-hand panels) for the duration.

<p>Group data–between-subject variation (mean concentrations, 95% confidence limits) of intact proinsulin (upper panel), insulin (middle panel) and the proinsulin:insulin ratio (lower panel) at collection (0h) and at the five time points thereafter, in samples kept at 4°C (left-hand panels) and at room temperature (right-hand panels) for the duration.</p

Additional file 2: Figure S1. of Phosphorylation of a splice variant of collapsin response mediator protein 2 in the nucleus of tumour cells links cyclin dependent kinase-5 to oncogenesis

A- GST-Tau was incubated with p35/CDK5 or p25/CDK5 and [γ-32P]-ATP for the times indicated, then subjected to SDS-PAGE prior to autoradiography (upper section). Tau bands were digested with Lys-C and phosphopeptides isolated and identified as described in Methods. A major phosphopeptide eluted with mass/charge ratio of 802.4331 corresponding to the peptide containing Ser235 (lower section), and a second doubly phosphorylated peptide corresponding to a peptide including Ser202 and Thr205 was also identified but at much lower abundance. The same result was obtained from two different phosphorylation reactions. B- GST-tau was phosphorylated as above but using non-radioactive ATP. Proteins were transferred to nitrocellulose after SDS-PAGE and probed with the indicated antibodies. Data is representative of two experiments. Figure S2. Co-expression of CDK5 complexes with CRMP2 and CRMP4. Hela cells were co-transfected with equal amounts of expression constructs for CDK5 catalytic subunit, p35 or p25 and FLAG-tagged CRMP2 (A and B) or CRMP4 (C and D) as indicated. Cells were lysed and protein expression and phosphorylation assessed by Western blot analysis (A and C). Quantification was performed on a Licor Odyssey (B and D) with data shown as mean ± S.E.M of three experiments in duplicate. t-test, *P < 0.05, **P < 0.01, ***P < 0.001. Figure S3. Co-expression of CDK5 complexes with tau. Hela cells were co-transfected with equal amounts of expression constructs for CDK5 catalytic subunit, p35 or p25 and tau. Cells were lysed and protein expression and phosphorylation assessed by (A) Western blot analysis. (B-D) Quantification was performed on a Licor Odyssey and the ratio between phospho-tau: total tau calculated for each phospho-tau antibody. Data shown as mean ± S.E.M. for three experiments performed in duplicate. t-test, *P < 0.05, **P < 0.01, ***P < 0.001. (PDF 2557 kb

Summary table.

<p>The study group is presented from the lower to the higher M-value, demonstrating clustering of signalling abnormalities stratified in ascending order according to the induction of signalling changes in response to insulin. Least potent induction is ranked as lowest L1 to L4; most potent induction is ranked as highest H1 to H4.</p

Fold activation of ERK by insulin according to body mass index or M value.

<p>(A) Body mass index (r = 0.73; p = 0.0009) or (B) M value (r = 0.52; p = 0.04).</p

Relationship of IRS1 expression with body mass index or M value.

<p>Relative IRS1 protein expression according to body mass index (A) or to M value (B) and fold increase in IRS1 expression according to body mass index (r = −0.36; p = 0.10) (C) or to M value (r = 0.27; p = 0.23) (D).</p

Relationship of PKB phosphorylation with body mass index or M value.

<p>Relative PKB phosphorylation according to body mass index (A) or to M value (B) and fold increase in PKB phosphorylation by insulin according to body mass index (r = −.38; p = 0.09) (C) or to M value (r = 0.4; p = 0.08) (D).</p

Representative Western blots.

<p>Body mass indices (BMI) are shown in parentheses and effects of fasting (−) or insulin (+).</p

Relationship between whole body insulin sensitivity and body mass index, lean body mass and percentage fat mass.

<p>Whole body insulin sensitivity (M value; mg/kg/min) and A) body mass index (kg/m²), B) lean body mass (kg) and C) percentage fat mass.</p

Relationship of ERK phosphorylation with body mass index or M value.

<p>Relative ERK phosphorylation according to body mass index (A) or to M value (B) and fold increase in ERK phosphorylation by insulin according to body mass index (r = 0.4; p = 0.07) (C) or to M value (r = 0.59; p = 0.08) (D).</p