Journal: Journal of Cellular and Molecular Medicine
Article Title: Tumour suppressor PTEN activity is differentially inducible by myo‐inositol phosphates
doi: 10.1111/jcmm.17699
Figure Lengend Snippet: (A, B) Effect of ITPP and BPBPP on PTEN phosphatase activity on PIP3 by measurement of PIP2 production. (A) Soluble PIP3 (5.0 × 10 −6 M) hydrolysis by PTEN (0.5 × 10 −6 M), performed in the presence of variable concentrations (0.1 × 10 −6 to 100.0 × 10 −6 M) of ITPP measured by anti‐PIP2 antibodies binding competition. (B) Soluble PIP3 (5.0 × 10 −6 M) hydrolysis by PTEN (0.5 × 10 −6 M) performed in the presence of variable concentrations (0.1 × 10 −6 to 100.0 × 10 −6 M) of BPBPP measured by anti‐PIP2 antibodies binding competition. The data are the means of three separate experiments in triplicate. The data are the means ± SD of three separate experiments in triplicate (* p < 0.05, ** p < 0.01). (C, D) Effect of ITPP and BPBPP on the activity of PI3‐Kinase on PIP2 by measuring PIP3 production. (C) Soluble PIP2 (5.0 × 10 −6 M) phosphorylation by PI3K (0.5 × 10 −6 M) was performed in the presence of variable concentrations (0.1 × 10 −6 to 100.0 × 10 −6 M) of ITPP measured by anti‐PIP3 antibodies binding competition. (D) Soluble PIP2 (5.0 × 10 −6 M) phosphorylation by PI3K (0.5 × 10 −6 M) performed in the presence of variable concentrations (0.1 × 10 −6 to 100.0 × 10 −6 M) of BPBPP measured by anti‐PIP3 antibodies binding competition. The data are the means of three separate experiments in triplicate.
Article Snippet: PTEN biochemical activity towards ITPP, BPBPP and IHP was assessed by an ELISA competition assay using the lipid Phosphatase Activity Assay to quantify PTEN activity (ELISA kit K‐4700, Echelon Biosciences, Salt Lake City, UT) following the manufacturer's protocol, designed to quantify the phosphatase activity of PTEN by the detection of the enzyme product, PI(4,5)P 2 .
Techniques: Activity Assay, Binding Assay