AFM / Confocal images and dataset

This dataset consist on AFM/confocal confinement at 3µm of HeLa Kyoto MYH9-eGFP / lifeact-mCherry.

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).

The following pharmacological inhibitors and chemical
compounds were used:10 µM ROCK-mediated contractility inhibitor Y-27632 (Y27) (ED Millipore),20 µM AACOCF3
(AA) inhibiting the nuclear envelope stretch-sensitive enzyme cPLA2 (Tocris
Bioscience),100 µM 2APB
blocking stretch-activated inositol triphosphate receptors (InsP3Rs) on the
ER/nuclear membranes (Tocris Bioscience).