Wes：阐述新冠病毒感染机制，推进抗病毒药物研发

比来的预印本杂志bioRxiv揭橥了德国波恩大学，夏里特大学，德国马克斯·普朗克神经病研究所等单元单子结合研究功效，阐述了SARS-CoV-2病毒能够按捺自噬。亚精胺，MK-2206，氯硝柳胺等感化于自噬的小分子化合物，能够作为抗病毒潜在治疗药物。

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Analysis of SARS-CoV-2-controlled autophagy reveals spermidine, MK-2206, and niclosamide as  putative antiviral therapeutics（bioRxiv preprint doi: https://doi.org/10.1101/2020.04.15.997254）

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--研究摘要--

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)对民众卫生和世界经济组成严重威胁，至今没有核准的特效药物或疫苗。代谢依靠的细胞途径的药理学调节，如自噬，削减了流传高致病性中东呼吸综合征(MERS)-CoV。（该单元单子之前研究，SKP2 Attenuates Autophagy Through Beclin1-ubiquitination and Its Inhibition Reduces MERS-Coronavirus Infection, Nat Commun, 10 (1), 5770 2019 Dec 18.）

本研究，作者发现SARS-CoV-2传染经由干扰多种代谢途径来限制自噬。体外靶向自噬诱导的化合物能够削减SARS-CoV-2在体外的释放。对自噬旌旗和代谢组学的深入剖析表明，SARS-CoV-2经由限制AMP-卵白激活激酶（AMPK)和mTORC1的激活，来削减糖酵解和卵白质翻译。传染还下调了自噬诱导的亚精胺，并促进AKT1/SKP2依靠途径自噬启动卵白Beclin-1(BECN1)降解。外源性给药亚精胺、AKT按捺剂MK-2206和不乱Beclin-1、抗螺旋肽药物氯硝柳胺离别按捺SARS-33CoV-2的释放达到85%、88%和99%。能够作为潜在的抗SARS-CoV-2的药物。

--Wes全主动定量Western Blot检测ATG14自然寡聚体--

ATG14是新冠病毒传染，按捺自噬旌旗通路中的要害卵白，其寡聚体降低的水平回响自噬被按捺的水平。

Wes全主动定量Western blot所有究竟为全膜数据，能够在单次实验中，同时检测卵白单体及其寡聚体，并定量转变。

Fig.2: SARS-CoV-2 hijacks autophagy signaling on multiple regulatory levels. VeroFM cells were infected with SARS-CoV-2 319 (MOI = 0.0005) and harvested at 8 h, 24 h, or 48 h post infection (h p.i.). Protein levels and phosphorylation status of selected autophagy-relevant proteins were determined by Western blotting. For analysis of ATG14 oligomers (bottom panel, left) cells were incubated with a chemical crosslinker (DSS) 2 h prior to cell harvest (for detailed protocol see methods section). Cell  extracts were separated and immunoblotted using Wes (ProteinSimple) capillary electrophoresis. Error bars in all panels denote standard error of mean derived from n = 3 biologically independent experiments. Tp < 0.1, *p < 0.05, ***p < 0.001  (one-way ANOVA; Bonferroni correction (post hoc)).