P70s6k

Manufactured by Cell Signaling Technology

Sourced in United States, China, Canada, United Kingdom, Germany

The P70S6K is a protein kinase that plays a key role in the regulation of cell growth, proliferation, and survival. It is a downstream effector of the PI3K/Akt signaling pathway and is involved in the phosphorylation of the ribosomal protein S6, which is important for protein synthesis and cell growth.

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Lab products found in correlation

Close spelling variants of this product

Anti-total p70S6K (5 citations) Phospho-p70S6K (103 citations) Anti-p-p70S6K (33 citations) P70S6K siRNA (2 citations) P-p70S6K-T421/S424 (3 citations) Anti-phospho-RPS6KB/p70S6K (Thr389) (2 citations) Phospho-Thr 389 p70S6K (3 citations) Rabbit anti-phosphor-p70S6K (Thr389) (2 citations) Anti-phospho-p70S6K-T389 (5 citations) Rabbit anti-phospho-p70S6K (3 citations)

353 protocols using p70s6k

Osthole Signaling Pathway Regulation

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Osthole (catalog number: O9265) was purchased from Sigma (St. Louis, MO, USA). Osthole was dissolved in DMSO to prepare a chemical stock for treatment. Antibodies against phosphorylated Akt (Ser⁴⁷³, catalog number: 4060), P70S6K (Thr⁴²¹/Ser⁴²⁴, catalog number: 9204), S6 (Ser²³⁵/Ser²³⁶, catalog number: 2211), ERK1/2 (Thr²⁰²/Tyr²⁰⁴, catalog number: 9101), p90RSK (Thr⁵⁷³, catalog number: 9346), JNK (Thr¹⁸³/Tyr¹⁸⁵, catalog number: 4668), total Akt (catalog number: 9272), P70S6K (catalog number: 9202), S6 (catalog number: 2217), ERK1/2 (catalog number: 4695), p90RSK (catalog number: 9335), JNK (catalog number: 9252), IRE1α (catalog number: 3294), eIF2α (catalog number: 5324), Bak (catalog number: 12105S), and Bax (catalog number: 2772) were purchased from Cell Signaling Technology (Beverly, MA, USA). Bcl-xL, p-Bcl-2, cleaved caspase 3 and cleaved caspase 9 were also purchased from cell Signaling Technology. Antibodies against GRP78 (catalog number: sc-13968), ATF6 α (catalog number: sc-166659), and α-tubulin (TUBA, catalog number: sc-32293) were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Inhibitors of ERK1/2 (U0126, catalog number: E1282) and JNK (SP600125, catalog number: E1305) were purchased from Enzo Life Sciences, Inc (Farmingdale, NY, USA), and a PI3K/Akt inhibitor (LY294002, catalog number: 9901) was purchased from Cell Signaling Technology, Inc.

Park W., Park S., Song G, & Lim W. (2019). Inhibitory Effects of Osthole on Human Breast Cancer Cell Progression via Induction of Cell Cycle Arrest, Mitochondrial Dysfunction, and ER Stress. Nutrients, 11(11), 2777.

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Hesperidin Modulation of Stress Signaling

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Hesperidin was purchased from Sigma-Aldrich and dissolved in dimethyl sulfoxide. Antibodies against phosphorylated AKT, P70S6K, ERK1/2, P90RSK, P38, eIF2α, and total AKT, P70S6K, ERK1/2, P90RSK, P38, IRE1α, and eIF2α were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against ATF6α was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). LY294002 was purchased from Cell Signaling Technology (Beverly, MA, USA). U0126 and SB203580 were purchased from Enzo Life Sciences (Farmingdale, NY, USA). Cisplatin and paclitaxel were purchased from Sigma-Aldrich (St.Louis, MO, USA).

Jeong S.A., Yang C., Song J., Song G., Jeong W, & Lim W. (2022). Hesperidin Suppresses the Proliferation of Prostate Cancer Cells by Inducing Oxidative Stress and Disrupting Ca2+ Homeostasis. Antioxidants, 11(9), 1633.

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Delphinidin Signaling Pathway Analysis

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Delphinidin was purchased from Indofine Chemical Company, Inc. (Hillsborough, NJ, USA). SB203580 and U0126 were purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA) and LY294002 was obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA). The antibodies against phosphorylated-AKT [serine (Ser)⁴⁷³] (catalog no. 4060), ERK1/2 [threonine (Thr)²⁰²/Tyr²⁰⁴, (catalog no. 9101; Cell Signaling Technology, Inc.)] c-Jun N-terminal kinase (JNK; Thr¹⁸³/Thr^185, catalog no. 4668), p38 (Thr¹⁸⁰/Thr¹⁸²; catalog no. 4511), ribosomal protein S6 kinase β-1 (P70S6K; Thr⁴²¹/Ser⁴²⁴; catalog no. 9204) and S6 (Ser^235/236, catalog no. 2211), and total AKT (catalog no. 9272), ERK1/2 (catalog no.4695), JNK (catalog no. 9252), p38 (catalog no. 9212), P70S6K (catalog no. 9202) and S6 (catalog no. 2217) were purchased from Cell Signaling Technology, Inc. All antibodies were diluted to 1:1,000 for western blot analysis. Cisplatin and paclitaxel were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany).

Lim W, & Song G. (2017). Inhibitory effects of delphinidin on the proliferation of ovarian cancer cells via PI3K/AKT and ERK 1/2 MAPK signal transduction. Oncology Letters, 14(1), 810-818.

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Western Blot Analysis of Signaling Proteins

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Phospho-specific anti-rabbit primary antibodies for P70s6k (Thr 421, S424), rpS6 (S235, S236), and 4E-BP1 (S65) (Cell Signaling, Danvers, MA) were visualized using chemiluminescent-labelled secondary antibodies (Amersham ECL Prime Western Blot Detector, GE Healthcare and Life Sciences, Pittsburgh, PA), and imaged with a Fluorchem SP system (Protein Simple Inc. Santa Clara, CA). Blots were then stripped for 20 minutes (glycine 15g, SDS 1g, Tween 20 10mL, QS ultra-pure water 1L, pH 2.2) before being reimaged to verify removal of all secondary and primary antibodies. Blots were then reprobed for total content of P70s6k, rps6, and 4E-BP1 (Cell Signaling, Danvers, MA).

Lane M., Herda T., Fry A., Cooper M., Andre M, & Gallagher P. (2017). Endocrine responses and acute mTOR pathway phosphorylation to resistance exercise with leucine and whey. Biology of Sport, 34(2), 197-203.

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Ipriflavone Inhibits mTOR Signaling

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Ipriflavone (7-isopropoxy-3-phenyl-4H-1-benzopyran-4-one) was purchased from TCI (Shanghai, China). AZD8055 was purchased from Selleckchem (Houston, TX, USA). Z-VAD-FMK was purchased from TOPSCIENCE (Shanghai, China). Aurintricarboxylic acid and BAX channel blocker were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). RPMI1640 medium and FBS were purchased from Biological Industries (Cromwell, CT, USA). Ham’s F12 medium was purchased from Lonza (Walkersville, MD, USA) and MEM/EBSS was purchased from Nanjing Keyjen Biotech (Nanjing, China). Active mTOR and p70S6K human recombinant proteins for kinase assays were purchased from SignalChem (Richmond, BC, Canada). Antibodies (1:1000) to detect phosphorylated AKT (S473), -mTOR (S2448), -mTOR (S2481), -EGFR (Y1068), -p70S6K (T389), -RSK(S380), -ERK1/2 (T202/Y204), total AKT, -mTOR, -EGFR, -p70S6K, -RSK, -ERK1/2, cyclin D3, p21, Cytochrome c, cleaved PARP and cleaved CASP3 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies to detect β‐actin (1:2000) were from Santa Cruz Biotechnology (Santa Cruz). Alanine aminotransferase Assay Kit and Aspartate aminotransferase Assay Kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Shi X., Zhang Y., Xie X., Pang M., Laster K., Li J., Ma X., Liu K., Dong Z, & Kim D.J. (2021). Ipriflavone Suppresses Growth of Esophageal Squamous Cell Carcinoma Through Inhibiting mTOR In Vitro and In Vivo. Frontiers in Oncology, 11, 648809.

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Investigating FLT3 Signaling Pathways

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MOLM14, MV4-11 cells were treated with DMSO, serially diluted A674563, TCS359 (1 μM), MK2206 (1 μM) for 4 hours. Cells were then washed in PBS and lysed in cell lysis buffer. FLT3, Phospho-FLT3 (Tyr589/591), AKT, Phospho-AKT Ser473, Phospho-AKT Thr308, GSK-3β, Phospho-GSK-3β (Ser9), Phospho-FoxO1 (Thr24), FoxO1, PRAS40, Phospho-PRAS40 (Thr246), STAT5, Phospho-STAT5 (Tyr694), NF-ΚB-P65, Phospho-NF-ΚB-P65 (Ser536), P70S6K, Phospho-P70S6K Thr389, 4EBP1, Phospho-4EBP1 (Thr37/46), ERK, Phospho-p44/42MAPK (Erk1/2) (Thr202/Tyr204), C-Myc and GAPDH antibodies (Cell Signaling Technology) were used for immunoblotting.
For FL addition experiment, MV4-11 cells were treated with FLT3 inhibitors in the presence of 10 ng/mL of FL (FLT3 Ligand, R&D Systems) or absence of FL for 2 hrs. Cells were then washed in PBS and lysed in cell lysis buffer. FLT3, Phospho-FLT3 (Tyr589/591), AKT, GSK-3β, Phospho-GSK-3β (Ser9), Phospho-FoxO1 (Thr24), FoxO1, STAT5, Phospho-STAT5 (Tyr694), P70S6K, Phospho-P70S6K (Thr389), 4EBP1, Phospho-4EBP1 (Thr37/46), C-Myc and GAPDH antibodies (Cell Signaling Technology) were used for immunoblotting.

Wang A., Wu H., Chen C., Hu C., Qi Z., Wang W., Yu K., Liu X., Zou F., Zhao Z., Wu J., Liu J., Liu F., Wang L., Stone R.M., Galinksy I.A., Griffin J.D., Zhang S., Weisberg E.L., Liu J, & Liu Q. (2016). Dual inhibition of AKT/FLT3-ITD by A674563 overcomes FLT3 ligand-induced drug resistance in FLT3-ITD positive AML. Oncotarget, 7(20), 29131-29142.

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Fraxetin Signaling Pathway Profiling

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Fraxetin (Cat No. 18224) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in dimethyl-sulfoxide (DMSO). The antibodies against phosphorylated extracellular-signal-regulated kinase (ERK)1/2 (Thr²⁰²/Tyr²⁰⁴, Cat No. 9101), JNK (Thr¹⁸³/Tyr¹⁸⁵, Cat No. 4668), P90RSK (Thr⁵⁷³, Cat No. 9346), P70S6K (Thr⁴²¹/Ser⁴²⁴, Cat No. 9204), S6 (Ser²³⁵/²³⁶, Cat No. 2211) and the total form of ERK1/2 (Cat No. 4695), JNK (Cat No. 9252), RSK1/RSK2/RSK3 (Cat No. 9355), P70S6K (Cat No. 9202), and S6 (Cat NO. 2217) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Song J., Ham J., Hong T., Song G, & Lim W. (2021). Fraxetin Suppresses Cell Proliferation and Induces Apoptosis through Mitochondria Dysfunction in Human Hepatocellular Carcinoma Cell Lines Huh7 and Hep3B. Pharmaceutics, 13(1), 112.

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Molecular Signaling Pathway Inhibitors

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Coumestrol, cisplatin and paclitaxel were purchased from Sigma-Aldrich, Inc. Rapamycin (mTOR inhibitor), SB203580 (P38 inhibitor), and U0126 (ERK1/2 inhibitor) were from Enzo Life Science (Farmingdale, NY, USA) and LY294002 (PI3K/AKT inhibitor) was from Cell Signaling Technology (Beverly, MA, USA). The antibodies against phosphorylated AKT (Ser 473 ), ERK1/2 (Thr 202 /Tyr 204 ), JNK1/2 (Thr 183 /Thr 185 ), p38 (Thr 180 /Thr 182 ), p70S6K (Thr 421 /Ser 424 ), p90RSK (Thr 573 ), and S6 (Ser 235/236 ) and total AKT, ERK1/2, JNK1/2, p38, p70S6K, p90RSK, and S6 were purchased from Cell Signaling Technology.

Lim W., Jeong W, & Song G. (2016). Coumestrol suppresses proliferation of ES2 human epithelial ovarian cancer cells. The Journal of endocrinology, 228(3).

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Autophagy and Apoptosis Pathway Analysis

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BCa cell lines were seeded in six-well plates, and after 24 h of culture, the cells were treated with NA at different dose and time. Western blotting was performed using specific antibodies against, Atg3, Atg5, Atg7, Atg12, beclin 1, LC3A, LC3B (Autophagy antibody sampler kit #4445, Cell Signaling), mTOR (#2972), pmTOR (#2974), p70S6K (#2708), phospho-p70S6K (#9234), BAX (#2772), BCL-2 (#2876), cleaved caspase-3 (#9661), cleaved caspase-9 (#9505), PARP (#9542) and cleaved PARP (#9541) (Cell Signaling), and β-actin (Santa Cruz Biotechnology, Dallas, TX, USA). Protein–antibody complexes were detected with the enhanced chemiluminescence method as described earlier (Suman et al, 2013b (link)).

Suman S., Das T.P., Reddy R., Nyakeriga A.M., Luevano J.E., Konwar D., Pahari P, & Damodaran C. (2014). The pro-apoptotic role of autophagy in breast cancer. British Journal of Cancer, 111(2), 309-317.

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Protein Quantification and Western Blotting Protocol

Cited 2 times

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Bicinchoninic acid (BCA) protein assay (Pierce Biotechnology, Rockford, IL, USA) was used to determine the protein content in each sample. Quantified each sample concentration to 60-80 µg and proteins were fractionated on SDS-PAGE, transferred onto Hybond enhanced chemiluminescence nitrocellulose membranes (Amersham, Little Chalfont, UK) and detected with antibodies against P-gp (GeneTex, Inc. Irvine, CA, USA), the mammalian target of rapamycin (mTOR) (Cell Signaling, Danvers, MA, USA), phosph-mTOR (Cell Signaling), protein kinase B (AKT) (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA), phosph-AKT (Santa Cruz Biotechnology, Inc.), p70s6K (Cell Signaling), phosph-p70s6K (Cell Signaling), caspase 3 (Cell Signaling), β-actin (Sigma Aldrich). Rabbit anti-mouse IgG-peroxidase antibody (Sigma Aldrich) and goat anti-rabbit IgG-peroxidase antibody (Sigma Aldrich) were used as the secondary antibody and protein-antibody complexes were visualized by enhanced chemiluminescence system. The Western blotting signals were quantified with ImageJ software (rsbweb.nih.gov/ij/) 14 (link), 15 (link).

Yang C.J., Chang W.W., Lin S.T., Chen M.C, & Lee C.H. (2018). Salmonella Overcomes Drug Resistance in Tumor through P-glycoprotein Downregulation. International Journal of Medical Sciences, 15(6), 574-579.

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