Plan apo vc 60x 1.4 oil objective

Cells were seeded in 35mm glass-bottom dishes (MatTek Corporation). Proximity ligation assays were performed using the Duolink In Situ Red Starter Kit Mouse/Rabbit (Millipore Sigma). In brief, cells were washed carefully and fixed in 4% paraformaldehyde. Cells were then permeabilized in 0.1% Triton-X/PBS before blocking. Primary antibodies were added overnight at 4 °C. PLA probes were added for 1 hour at 37 °C before ligation and amplification. After washing and staining cell nuclei with DAPI solution (1:1000 in PBS, stock 1 mg/mL, Thermo Fisher Scientific), cells were imaged at a Yokogawa CSU22 spinning disk confocal microscope using a Plan Apo VC 60X/ 1.4 Oil objective (Nikon Imaging Center, UCSF). Image analysis was done via Fiji ImageJ software 58

Cells were seeded in 35mm glass-bottom dishes (MatTek Corporation) or µClear 96-well imaging plates (Greiner Bio-One). At the time of harvest, cells were washed carefully and fixed in 4% paraformaldehyde. Cells were then permeabilized in 0.1% Triton-X/PBS and blocked in 5% bovine serum albumin (BSA) / 0.1% Triton-X/PBS. Primary antibody was diluted in 1% BSA / 0.1% Triton-X/PBS and incubated overnight at 4 °C. After washing, secondary antibodies were diluted in 1% BSA / 0.1% Triton-X/PBS and added for 1 hour at room temperature.
Where indicated, actin filaments were stained with rhodamine-phalloidin (Thermo Fisher Scientific) as described by the manufacturer. After secondary antibody staining, cells were washed and then stained with DAPI solution (1:1000 in PBS, stock 1 mg/mL, Thermo Fisher Scientific). For endogenously tagged YAP-mNeonGreen2 cells, permeabilization, blocking as well as primary and secondary antibody staining were omitted. Cells were imaged at a Yokogawa CSU22 spinning disk confocal microscope using a Plan Apo VC 60X/ 1.4 Oil objective (Nikon Imaging Center, UCSF). Image analysis was done via Fiji ImageJ software 58 (link) . Quantification of relative integrated density for nuclear levels was performed by automated analysis quantifying the intensity for the protein of interest per nuclei.