Tfa2c

Trophoblast organoids were washed using the cell recovery solution (Corning, USA) and then fixed in 4% paraformaldehyde for 30 min. After permeation and blocking, primary antibodies against human GATA3 (1:100, Abcam, UK), TFA2A (1:100, Abcam, UK) and TFA2C (1:100, Abcam, UK) were added and incubated at 4℃ overnight. On the next day, the organoids were sequentially incubated with fluorescence-labeled rabbit or mouse secondary antibody. Phalloidine and DAPI were treated orderly. Imaging of the stained organoids was carried out by a ZEISS LSM 700 Confocal Laser Scanning Microscope and the ZEN software.

The organoids were washed with cell recovery solution (Corning, USA) before being fixed for 30 min in 4% paraformaldehyde. After permeation and blocking, primary antibodies against human GATA3 (1,100, Abcam, UK), TFA2A (1,100, Abcam, UK), TFA2C (1,100, Abcam, UK), beta-hCG (1,500, Abcam, UK), E-cadherin (1,250, Abcam, UK), HLA-G (1,50, Abcam, UK), and PAPPA2 (1,100, Abcam, UK) were added and incubated at 4°C overnight. The following day, the organoids were sequentially incubated with fluorescence-labeled secondary rabbit or mouse antibody. Phalloidin and DAPI were added accordingly. Subsequently, the stained organoids were imaged using a ZEISS LSM 700 Confocal Laser Scanning Microscope and ZEN imaging software.