AFM / Confocal images and dataset

This dataset consist on AFM/confocal confinement at 3µm of HeLa Kyoto MYH9-eGFP / lifeact-mCherry.<br> Dishes with cells were mounted in a dish heater (JPK Instruments) and kept at 37 °C under an inverted light microscope (Axio Observer.Z1; Zeiss) equipped with a confocal microscope unit (LSM 700; Zeiss) and the atomic force microscope (AFM) head (CellHesion 200; JPK Instruments). Focused ion beam (FIB)-sculpted, flat silicon microcantilevers were processed and calibrated as described in (Cattin et al., 2015). The microcantilevers were fixed on a standard JPK glass block and mounted in the AFM head. The cantilever was lowered on the cell to a preset height with a constant speed of 0.5 μm·s-1, and the resulting varying force and cantilever height were recorded over time. At the same time, differential interference contrast (DIC) and fluorescence images at the confined cell's midplane were recorded every 5 seconds using a 63× water immersion objective. All microscopy equipment was placed, and experiments were carried out in a custom-made isolation box at 37 °C (The Cube; Life Imaging Services).<br> The following pharmacological inhibitors and chemical compounds were used:10 µM ROCK-mediated contractility inhibitor <b>Y-27632</b> (Y27) (ED Millipore),20 µM <b>AACOCF3</b> (AA) inhibiting the nuclear envelope stretch-sensitive enzyme cPLA2 (Tocris Bioscience),100 µM <b>2APB</b> blocking stretch-activated inositol triphosphate receptors (InsP3Rs) on the ER/nuclear membranes (Tocris Bioscience).本数据集包含经3μm约束的原子力显微镜(atomic force microscope, AFM)/共聚焦成像数据，样本为转染MYH9-eGFP与lifeact-mCherry的HeLa Kyoto细胞。 将接种细胞的培养皿置于培养皿加热器（JPK Instruments）中，维持37℃温度，放置于配备共聚焦成像单元（LSM 700；蔡司）与AFM探头（CellHesion 200；JPK Instruments）的倒置光学显微镜（Axio Observer.Z1；蔡司）下。聚焦离子束(focused ion beam, FIB)刻蚀的平面硅微悬臂梁按照Cattin等（2015）的研究方法进行制备与校准。将微悬臂梁固定于标准JPK玻璃基座后，安装至AFM探头中。以0.5 μm·s⁻¹的恒定速率将悬臂梁降至细胞上方预设高度，随时间同步记录悬臂梁所受变力与自身高度的变化。同时，采用63倍水浸物镜，每5秒记录一次受约束细胞中央平面的微分干涉差(differential interference contrast, DIC)成像与荧光图像。所有显微成像设备均安置于定制恒温隔离箱（The Cube；Life Imaging Services）内，实验全程维持37℃环境。 本实验使用以下药理学抑制剂与化学试剂：10 μM ROCK介导收缩力抑制剂<b>Y-27632</b>（简称Y27，ED Millipore）；20 μM <b>AACOCF3</b>（简称AA），可抑制核被膜牵张敏感性酶cPLA2（Tocris Bioscience）；100 μM <b>2APB</b>，可阻断内质网/核膜上的牵张激活型三磷酸肌醇受体(inositol triphosphate receptors, InsP3Rs)（Tocris Bioscience）。