AFM / Confocal 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).