| 货号 | 6286S |
| 描述 | SignalSilence® Atg14 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Atg14 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis. |
| 反应种属 | Human |
| 应用 | TFN |
| 供应商 | CST |
| 背景 | Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but is also associated with a number of physiological processes including development, differentiation, neurodegeneration, infection and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and is directed by a number of autophagy-related (Atg) genes. These proteins are involved in the formation of autophagosomes, cytoplasmic vacuoles that are delivered to lysosomes for degradation. The class III type phosphoinositide 3-kinase (PI3K) Vps34 regulates vacuolar trafficking and autophagy (4,5). Multiple proteins associate with Vsp34, including p105/Vsp15, Beclin-1, UVRAG, Atg14, and Rubicon, to determine Vsp34 function (6-12). Atg14 and Rubicon were identified based on their ability to bind to Beclin-1 and participate in unique complexes with opposing functions (9-12). Rubicon, which localizes to the endosome and lysosome, inhibits Vps34 lipid kinase activity; knockdown of Rubicon enhances autophagy and endocytic trafficking (11,12). In contrast, Atg14 localizes to autophagosomes, isolation membranes and ER, and can enhance Vps34 activity. Knockdown of Atg14 inhibits starvation-induced autophagy (11,12).自噬是自噬溶酶体降解其所包裹的细胞质内容物的一个分解代谢过程。通常情况下，自噬在营养匮乏的时候被激活，但它还与许多生理过程相关，包括生长、分化、神经退行性疾病、感染和癌症。自噬的分子机制主要在酵母中发现的，并称之为自噬相关基因。这些蛋白与自噬体形成有关，自噬体就是被送往溶酶体进行降解的胞浆空泡。III型磷酸肌醇3激酶（PI3K）Vps34调节空泡的运输和自噬（4,5）。许多蛋白包括p105/Vsp15，Beclin-1，UVRAG，Atg14和Rubicon与Vsp34一起决定Vsp34的功能（6-12）。Atg14和Rubicon能够与Beclin-1结合，并且参与具有相对功能的独特复合物的形成（9-12）。Rubicon主要集中于内涵体和溶酶体，能够抑制Vps34脂质激酶活性；Rubicon的下调可以增强自噬和内吞作用（11,12）。相反，Atg14集中于自噬体，隔离膜和内质网，能够增强Vps34活性；Atg14的下调可以抑制饥饿诱导的自噬(11,12)。 |
| 存放说明 | -20C |
| 参考文献 | Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88. Corvera, S. (2001) Traffic 2, 859-66. Yan, Y. and Backer, J.M. (2007) Biochem Soc Trans 35, 239-41. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34. Zeng, X. et al. (2006) J Cell Sci 119, 259-70. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96. |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Atg14 siRNA I (+) or SignalSilence® Atg14 siRNA II #6287 (+), using Atg14 Antibody #5504 (upper) or β-Tubulin (9F3) Rabbit mAb #2128 (lower). The Atg14 Antibody confirms silencing of Atg14 expression, while the β-Tubulin (9F3) Rabbit mAb is used as a loading control.Western blot分析HeLa细胞的提取物，用100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)，SignalSilence® Atg14 siRNA I (+) 或SignalSilence® Atg14 siRNA II #6287 (+)转染，使用的抗体是：Atg14 Antibody #5504（上图）， β-Tubulin (9F3) Rabbit mAb #2128 （下图）。在以 β-Tubulin (9F3) Rabbit mAb作为加样对照时，Atg14 Antibody 证实了Atg14 表达的沉默。 |
