Abstract：
Recent evidence has shown that uncoating and reverse transcription precede nuclear import. These recent breakthroughs have been made possible through the development of innovative biochemical and imaging techniques. This method outlines the biochemical assay used for detecting the presence of the HIV-1 core in the nuclear compartment. In this procedure, human cells are infected with HIV-1NL4-3, with or without the inclusion of PF74, a small molecule that inhibits core entry into the nuclear compartment. Subsequently, cells are separated into cytosolic and nuclear fractions. To assess whether the capsid protein has reached the nuclear compartment, cytosolic and nuclear fractions are subjected to Western blot analysis, utilizing antibodies specific to the HIV-1 capsid protein p24. To validate the true origin of these fractions, Western blot analysis employing antibodies against cytosolic and nuclear markers are also performed. In summary, this assay provides a reliable and efficient means to detect the presence of the HIV-1 capsid protein in the nucleus during infection under various conditions.
摘要：
最近的证据表明，脱涂层和逆转录先于核导入。通过创新的生化和成像技术的发展，这些最近的突破已经成为可能。该方法概述了用于检测核室中HIV-1核心存在的生化测定。在这个过程中，人类细胞感染HIV-1NL4-3，有或没有包含PF74，这是一种抑制核心进入核室的小分子。随后，细胞被分离为胞质和核部分。为了评估衣壳蛋白是否已经到达核室，胞质和核部分进行蛋白质印迹分析，利用特异于HIV-1衣壳蛋白p24的抗体。为了验证这些分数的真正来源，还进行了使用针对胞质和核标记的抗体的Western印迹分析。总之,该测定提供了一种可靠和有效的方法来检测在各种条件下感染期间细胞核中HIV-1衣壳蛋白的存在。
