Figure 1
Stage dependent expression of the different PI3K Type IA catalytic isoforms. A) Protein levels of the different PI3K Type IA catalytical subunits were analyzed with western blot using two sets of primary neuroblastoma tumors. Abbreviations: AG, adrenal gland; MNA, MYCN amplification B) Relative quantification (RQ) of p110α, β and δ protein levels in primary tumors. Boxplot explanation; upper hinge of the box, 75th percentile; lower hinge of the box, 25th percentile; thick horizontal line within box, median. The whiskers are indicating range, open circles represent outliers while asterisks represent extremes. C) Western blot of cell lines with different origin show that cell lines of neuroblastic origin has high levels of p110α and low levels of p110β compared to non neuroblastic cell lines. Grading of MNA; no gain or amplification (−), gain (+), amplified (++).
Figure 2
Stage dependent expression of the PI3K Type IA regulatory isoforms. A) Protein levels of the regulatory subunits of Type IA PI3K: p85α/β, p55α, p50α and p55γ were analyzed with western blot using two sets of primary neuroblastoma tumors. Abbreviations: AG, adrenal gland; B) Relative quantification (RQ) of p85α/β, p55α, p50α and p55γ protein levels using Gapdh as endogenous control. Boxplot explanation; upper hinge of the box, 75th percentile; lower hinge of the box, 25th percentile; thick horizontal line within box, median. The whiskers are indicating range, open circles represent outliers while asterisks represent extremes. C) Protein levels of regulatory subunits in neuroblastoma cell lines (to the left), and cell lines of other origins (to the right).
Figure 3
Phosphorylated Akt and correlation to PTEN. A) Western blot of primary neuroblastoma tumors showing levels of p-Akt/PKB (S473), p-Akt/PKB (T308), Akt/PKB and PTEN. AG = adrenal gland. B) Levels of p-Akt/PKB (T308), p-Akt/PKB (S473) and PTEN in stage 1–2 (st 1–2) and stage 4 (st 4) neuroblastoma tumors. Relative quantification (RQ) was calculated using GAPDH as endogenous control. C) Correlation plot using Spearman Rho showing that levels of phosphorylated Akt/PKB (T308) are inversely correlated with protein levels of PTEN.
Table 1
Clinical data
| Patient | INSS | INRG | Age at diagnosis (months) | Follow up or outcome change (months) | Outcome | 1p loss | MNA | 11q loss | Origin |
|---|---|---|---|---|---|---|---|---|---|
| 39R1 | 4 | M | 22 | 29 | NED | pos | pos | neg | Abdominal |
Abbreviations: INSS, International neuroblastoma staging system; INRG, International neuroblastoma risk group; MNA, MYCN amplification; NA, Information not available; L, Localized; M, Metastasized; NED, No evidence of disease; DOD, Dead of disease; DSC, Dead by surgical complications; pos, Positive; neg, Negative.