Extended Data Fig. 8: The desaturase SCD1 contributes to cytoskeleton organization and apical domain formation in eight-cell-to-blastocyst development.

(a) Immunofluorescence of K18 and CDX2 showing their location in 3.5 d.p.c blastocyst embryos treated with DMSO-, CAY10566- or CAY10566 plus oleic acid; the images shown are representative of one (for K18) and two (for CDX2) independent experiments. (b-d) Quantification of fluorescence intensity of the cell-cell boundary (b), cell number per embryo (c) and CDX2 intensity (d) of 3.5 d.p.c blastocyst embryos treated with DMSO-, CAY10566- or CAY10566 plus oleic acid. Each dot represents one blastocyst, Data (b, c) are from n = 5 for DMSO, n = 10 for CAY10566 and n = 6 for CAY10566 + OA-treated blastocysts, the experiment was performed for one time; data (d) are from n = 13 for DMSO, n = 20 for CAY10566 and n = 15 for CAY10566 + OA-treated blastocysts, and the experiments were repeated for two times. Data are presented as the mean +/-s.e.m. Statistical significance was determined by two-tailed unpaired t test. Center line, median; box, 25th and 75th percentiles, whiskers, 1.5xIQR. (e) Immunofluorescence of K18 and CDX2 of 3.5 d.p.c. blastocyst embryos treated with NC, siScd1 or siScd1 plus oleic acid. The images shown are representative of two (for siScd1) and one (for siScd1 + OA) independent experiments. (f, g) Quantification of fluorescence intensity of the cell-cell boundary (f), cell number per embryo (g) of 3.5 d.p.c blastocyst embryos treated with NC, siScd1 or siScd1 plus oleic acid. Each dot represents one blastocyst. Data are from n = 17 for siNC, n = 19 for siScd1 and n = 5 for siScd1 + OA-treated blastocysts, the experiments were repeated for two (for siScd1) and one (for siScd1 + OA) times. Data are presented as the mean +/-s.e.m. Statistical significance was determined by two-tailed unpaired t test. Center line, median; box, 25th and 75th percentiles, whiskers, 1.5xIQR.

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