Anti gata6

Mouse ESCs were cultured in 8-well slides (Ibidi). ESC immunostaining was carried out as previously described in Canham et al., 2010 (link). The following antibodies were used: anti-NANOG (eBioscience, 14-5761, 1:200), anti-OCT4 (Santa Cruz, sc-5279, 1:200), and anti-GATA6 (Cell Signalling Technologies, 5851, 1:1600). Secondary antibodies used are from the Alexa Fluor series (Molecular Probes, Thermo Fisher). mESCs were imaged using a Deltavision Widefield Screening microscope.

hCMPCs were transfected with miR-10a mimics or inhibitor for 48 h. Then, the cells were harvested and lysed with RIPA buffer (Biotime, China) for protein extraction. Proteinase inhibitors were used to minimize protein decomposition. Western blotting was performed using NuPAGE 4%–12% Bis-Tris Gel (NP0335BOX, Invitrogen, USA). The blots were incubated with primary antibodies anti-GATA6 (5851, Cell Signal Technology, USA), anti-caspase 3 (Cell Signaling, USA) and anti-GAPDH (Cell Signaling, USA) at 4°C overnight to detect the relative protein expression levels, and a fluorescent secondary antibody (072-07-15-06, KPL, USA) was applied to label the primary antibody. The labeled bands were visualized with Oddesey (Li-Cor).