Journal: PLoS ONE

Article Title: TGFβ-Stimulated MicroRNA-21 Utilizes PTEN to Orchestrate AKT/mTORC1 Signaling for Mesangial Cell Hypertrophy and Matrix Expansion

doi: 10.1371/journal.pone.0042316

Figure Lengend Snippet: Mesangial cells were transfected with miR-21 Sponge or vector. The serum-starved cells were incubated with 2 ng/ml TGFβ for 24 hours. The cell lysates were immunoblotted with phospho-S6 kinase (Thr-389), S6 kinase (panel A), phospho-mTOR (Ser-2448), mTOR (panel B), phospho-4EBP-1 (Thr-34/46), phospho-4EBP-1 (Ser-65) and 4EBP-1 (panel C) antibodies as indicated.

Article Snippet: Phospho-Akt (Ser473), phospho-Akt (thr-308), Akt, phospho-S6 kinase, S6 kinase, phospho-4EBP-1 (Thr-37/46) phospho-4EBP-1 (Ser-65) 4EBP-1, phospho-GSK3β, GSK3β, phospho-tuberin (Thr 1462), tuberin, phospho-PRAS40 (Thr 246), PRAS40, phospho-mTOR (Ser-2448) and mTOR antibodies were purchased from Cell Signaling, Boston, MA. siRNA pool of three oligonucleotides against PTEN mRNA, collagen II (α2) and PTEN antibodies were obtained from Santacruz, Delaware, CA.

Techniques: Transfection, Plasmid Preparation, Incubation

Journal: Oncoscience

Article Title: Genistein exerts anti-leukemic effects on genetically different acute myeloid leukemia cell lines by inhibiting protein synthesis and cell proliferation while inducing apoptosis – molecular insights from an iTRAQ™ quantitative proteomics study

doi:

Figure Lengend Snippet: A) Protein synthesis was analyzed using metabolic labelling, and there is an immediate and consistent drop in the protein synthesis rates in MV4-11 cells. However, a delayed effect is observed in the HL-60 cells, where significant decrease in protein synthesis was observed after 72 h of genistein treatment. The data shown is the mean of a triplicate of experiments. * indicates p<0.05; *** indicates p <0.001. Error bars represent standard deviation. B) Western blot analysis of mTOR pathway, which is the key pathway in regulating protein synthesis and is reported to be regulated from our iTRAQ analysis. Our result shows the reduced phosphorylation of the protein 4EBP-1, where the unphosphorylated form binds to eIF4E protein and inhibits its translational activity. This results in the decreased protein synthesis, in accordance to our protein synthesis assay.

Article Snippet: Antibodies against p-FLT3, p-4EBP-1 and 4EBP-1 were obtained from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Metabolic Labelling, Standard Deviation, Western Blot, Activity Assay

Journal: Oncoscience

Article Title: Genistein exerts anti-leukemic effects on genetically different acute myeloid leukemia cell lines by inhibiting protein synthesis and cell proliferation while inducing apoptosis – molecular insights from an iTRAQ™ quantitative proteomics study

doi:

Figure Lengend Snippet: A) Protein synthesis was analyzed using metabolic labelling, and there is an immediate and consistent drop in the protein synthesis rates in MV4-11 cells. However, a delayed effect is observed in the HL-60 cells, where significant decrease in protein synthesis was observed after 72 h of genistein treatment. The data shown is the mean of a triplicate of experiments. * indicates p<0.05; *** indicates p <0.001. Error bars represent standard deviation. B) Western blot analysis of mTOR pathway, which is the key pathway in regulating protein synthesis and is reported to be regulated from our iTRAQ analysis. Our result shows the reduced phosphorylation of the protein 4EBP-1, where the unphosphorylated form binds to eIF4E protein and inhibits its translational activity. This results in the decreased protein synthesis, in accordance to our protein synthesis assay.

Article Snippet: Antibodies against p-FLT3, p-4EBP-1 and 4EBP-1 were obtained from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Metabolic Labelling, Standard Deviation, Western Blot, Activity Assay