Ab110239

Cells were seeded onto Matrigel-coated PAGs and glass coverslips at a density of 7-9×10 6 in supplemented media. Media was aspirated and adherent cells were washed with 1x PBS, xed with 4 % paraformaldehyde (PFA, Sigma-Aldrich) (v/v) for 10 minutes at room temperature, then washed three times with 1x PBS and permeabilised for ve minutes in permeabilising solution -0.2 % Triton X-100 (v/v) (Sigma-Aldrich) in wash buffer (0.5 % bovine serum albumin (BSA, ThermoFisher, (w/v) in PBS).Permeabilising solution was aspirated, and the cells were washed three times with wash buffer. For TAZ staining, samples were rst blocked for 1 hour at room temperature with 5 % BSA in normal serum (10 % Donkey serum (sigma-Aldrich) (v/v) in 1x PBS). Cells were then incubated with a primary antibody targeting CTGF (1:1000, Ab6692, Abcam) or TAZ (1:200; Ab110239; Abcam) for 1 hour at room temperature followed by washing three times with wash buffer and incubation with uorescently-tagged secondary antibody (Abcam) diluted at 1:1000 in normal serum for 1 hour at room temperature. Cells were counter-stained with DAPI (Sigma-Aldrich) and uorescently-tagged phalloidin (ThermoFisher) using standard protocols and washed a nal three times with wash buffer and twice with distilled water before mounting onto microscope slides.