Rabbit anti gata6

Cells were grown on μ‐slides (Ibidi) and fixed in 4% paraformaldehyde (PFA) in D‐PBS (without Ca²⁺ and Mg²⁺) for 10 min at RT followed by PFA inactivation in 300 mM glycine in D‐PBS (5 min, RT) and a wash in D‐PBS. Cells were permeabilized with 1% (v/v) Triton X‐100, 0.2% (w/v) SDS, 10 mg/ml BSA in D‐PBS (1 h, RT) and incubated with primary antibodies overnight at 4°C in 50 mg/ml BSA in TNT (100 mM Tris–Cl (pH 7.5), 150 mM NaCl, and 0.1% (v/v) Tween‐20). The following antibodies and dilutions were used: rabbit anti‐TUJ1 (Cell Signaling, 5568) at 1:600; rabbit anti‐DESMIN (Cell Signaling, 5332) at 1:300; rabbit anti‐GATA6 (Cell Signaling, 5851) at 1:1,600. An anti‐rabbit IgG (H+L) F(ab′)₂ Fragment Alexa Fluor 647 Conjugate (Cell Signaling, 4414) served as secondary antibody and was allowed to incubate for 2 h at RT in 50 mg/ml BSA in TNT. Nuclei were visualized using a constitutive nuclear marker (a stably integrated CAG::H2B‐mCherry‐BGHpA plasmid). Confocal images were acquired on an inverted SP8 confocal microscope (Leica) equipped with a 40× PL Apo 1.1 W objective. TUJ1 immunostaining for quantification by flow cytometry was performed under the same conditions except that the starting material was a single‐cell suspension.