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![ALM inhibits HIF-1 alpha translation by down-regulating mTOR pathway. (A) Hep3B and PC3 cells were exposed to vehicle or the hypoxia mimics dimethyloxalylglycine (DMOG), cobalt chloride (CoCl2), desferrioxamine (DFX) or MG132 in the presence of ALM at indicated concentrations or vehicle for 24 h and whole cell lysates were subjected to Western blot. (B) Hep3B cells were pretreated for 4 h with vehicle (Vhc), 25 nM rapamycin (Rapa), 20 μg/mL cycloheximide (Chx), or 200 nM ALM, [35S] methionine/cysteine was added for 1 h followed by cell lysis, HIF-1 alpha immunoprecipitation (IP), SDS/PAGE, and autoradiography. (C–E) PC3 cells were cultured at 20% or 1% O2 for 24 h in the presence of ALM (at the indicated concentrations) and whole cell lysates were subjected to for: (C) p-P70, p-Akt, p-mTOR (2448/2481), mTOR, (D) p-4E-BP1, (E) p-RPS6, HIF-1 alpha , or beta -actin. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31083403), licensed under a CC-BY license. Not internally tested by R&D Systems.](http://images.novusbio.com/fullsize/af3918_phospho-rps6-s235-s236-affinity-purified-polyclonal-ab-41202412403591.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Thymus stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0011.jpg "Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Thymus stained with anti-S6 Ribosomal (Phospho S235/236) antibody.")
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Breast Carcinoma stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0001.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Breast Ductal Cell Carcinoma stained with anti-S6 Ribosomal (phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0002.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Bladder Transitional Cell Carcinoma stained with anti-S6 Ribosomal (Phospho S235/236)antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0003.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Colon Adenocarcinoma stained with anti-S6 Ribosomal (Phospho S235/236)antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0004.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Hepatocellular Cell Carcinoma stained with anti-S6 Ribosomal (Phospho S235/236)antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0005.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Lung Adenocarcinoma stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0006.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Lung Squamous Cell Carcinoma stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0007.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Renal Cell Carcinoma stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0008.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Stomach Adenocarcinoma stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0009.jpg)
![Immunohistochemistry-Paraffin: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody (SP50) [NBP2-12478] - Human Tonsil stained with anti-S6 Ribosomal (Phospho S235/236) antibody.](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-p-Ser235-p-Ser236-Antibody-SP50-Immunohistochemistry-Paraffin-NBP2-12478-img0010.jpg)
![ELISA Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] [Biotin]](http://images.novusbio.com/fullsize2/dyc3918-2_phospho-rps6-s235-s236-duoset-ic-elisa-2-plate-elisa-169202094547.jpg "ELISA Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] [Biotin]")
![Western Blot: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody [NB100-1553] - Detection of Human Phospho RPS6 (S235/S236) by Western Blot. Samples: Whole cell lysate (35 mcg) from HeLa cells. Treatments consisted of growth in serum containing media (lane 1) or cultured for 24 hours in serum free media (lanes 2 to 5). At 24h, serum was replaced for 1 h (lanes 2 and 3) or serum starvation was continued (lanes 4 and 5) without rapamycin (lanes 3 and 5) with 50nM Rapamcycin (lanes 2 and 4). Antibodies: Affinity purified rabbit anti-phospho RPS6 (S235/S236) antibody NB100-1553 used at 0.04 mcg/ml (upper panel). The blot was stripped and re-blotted for total RPS6 using affinity purified rabbit anti-RPS6 antibody NB100-1595 at 0.04 mcg/ml. Detection: Chemiluminescence with exposure times of 10 seconds (upper panel) or 30 seconds (lower panel).](http://images.novusbio.com/fullsize/Ribosomal-Protein-S6-RPS6-[p-Ser235--p-Ser236]-Antibody-Western-Blot-NB100-1553-img0002.jpg "Western Blot: Ribosomal Protein S6/RPS6 [p Ser235, p Ser236] Antibody [NB100-1553] - Detection of Human Phospho RPS6 (S235/S236) by Western Blot. Samples: Whole cell lysate (35 mcg) from HeLa cells. Treatments consisted of growth in serum containing media (lane 1) or cultured for 24 hours in serum free media (lanes 2 to 5). At 24h, serum was replaced for 1 h (lanes 2 and 3) or serum starvation was continued (lanes 4 and 5) without rapamycin (lanes 3 and 5) with 50nM Rapamcycin (lanes 2 and 4). Antibodies: Affinity purified rabbit anti-phospho RPS6 (S235/S236) antibody NB100-1553 used at 0.04 mcg/ml (upper panel). The blot was stripped and re-blotted for total RPS6 using affinity purified rabbit anti-RPS6 antibody NB100-1595 at 0.04 mcg/ml. Detection: Chemiluminescence with exposure times of 10 seconds (upper panel) or 30 seconds (lower panel).")