Donkey anti goat igg alexa fluor plus 488

Cells were fixed for 10 min in 3.7% paraformaldehyde at RT, permeabilized with 0.1% Triton X-100 for 15 minutes at RT and blocked in PBS with 5% BSA for 1 hour at RT. Cells were incubated with primary antibodies (anti-GATA6 (AF1700, R&D systems, 8 μl/ml) or anti-SOX1 (AF3369, R&D systems, 8 μl/ml)) in blocking solution overnight at 4°C, rinsed and incubated with secondary antibodies (Fig. 2g: donkey anti-goat IgG Alexa Fluor Plus 488 (A32814, Invitrogen, 1 μl/ml); Extended Data Fig. 5c: donkey anti-goat IgG Alexa Fluor Plus 594 (A32758, Invitrogen, 1 μl/ml)) in blocking solution for 30 minutes at RT. Nuclei were stained with DAPI (Sigma, 28718-90-3) and mounting with anti-fading mounting medium (Life Technologies, P10144).