Antibody:	anti-Cingulin
Detection system:	Secondary antibody
Assay notes:	For whole mount specimens, the fixative was 0.5% formaldehyde for 20 minutes. For sectioned specimens, the fixative was 1% formaldehyde.

Structure	Level	Pattern	Image	Note
TS28: oocyte	Present	Regionally restricted	1A , 1B	Expression was detected in the cytoplasmic face of the oolemma in 100% of oocytes. In the cytocortex, the microvillous domain was stained, but staining was absent or poor in the smooth membrane domain overlying the metaphase II spindle.

Structure	Level	Pattern	Image	Note
TS28: oocyte	Present	Regionally restricted	1D , 1C	Expression was detected in the oocyte cortex in 100% of the oocytes. Staining was not detected in most cumulus cells except within the corona radiata weakly at the site of contact with the oocyte.

Structure	Level	Pattern	Image	Note
TS28: oocyte	Present	Regionally restricted	1E , 1F	Expression was detected in the oocyte cortex in 100% of the oocytes. Staining was not detected in most cumulus cells except within the corona radiata weakly at the site of contact with the oocyte.

Structure	Level	Pattern	Image	Note
TS1: 1-cell stage embryo	Present	Regionally restricted		Expression was detected in the cytoplasmic face of the oolemma of 100% of the fertilized eggs.

Structure	Level	Pattern	Image	Note
TS2: 2-cell stage embryo	Present	Regionally restricted		Expression was detected in the cytocortex either uniformly or preferentially on the outer surface of 69% of embryos.

Structure	Level	Pattern	Image	Note
TS3: 4-cell stage embryo	Present	Regionally restricted	2A	Expression was detected in the cytocortex either uniformly or preferentially on the outer surface of 78% of embryos.

Structure	Level	Pattern	Image	Note
TS3: 8-cell stage embryo	Present	Regionally restricted		Expression in the cytocortex was detected in the apical microvillous pole on the outer surface of each blastomere in 57% of the embryos but absent from the nascent apicolateral tight junctions site.

Structure	Level	Pattern	Image	Note
TS3: compacted morula	Present	Regionally restricted	2B	Expression in the cytocortex was detected in the apical microvillous pole on the outer surface of each blastomere in 85% of the embryos but absent from the nascent apicolateral tight junctions site.

Structure	Level	Pattern	Image	Note
TS3: compacted morula	Present	Regionally restricted	2C	Expression in the cytocortex was in the apical microvillous pole in outer blastomeres of the trophectoderm lineage of 67% of embryos. Faint zonular or discontinuous staining was in the forming tight junction sites of each blastomere in 75% of embryos.

Structure	Level	Pattern	Image	Note
TS4: compacted morula	Present	Regionally restricted		Expression was absent from the inner cell mass and the cytocortex, but was present in the tight junctions in the trophectoderm in 100% of embryos and in punctate cytoplasmic foci surrounding the trophectoderm nuclei in 70% of embryos.