Fig. 3

IL6 supplementation preferentially increases PE cell numbers. Blastocysts were cultured in SOF-BE1 supplemented with either 0 or 100 ng/ml IL6 for various periods of time, then immuno-staining for PE (GATA6⁺), EPI (NANOG⁺) and TE (CDX2⁺) was completed. All NANOG⁺/GATA6⁺ dual-positive cells were considered UN cells. Panel a: IL6 supplementation from days 5 to 8 (n = 28–29 blastocysts/group over 3 replicate studies). Panel b: IL6 supplementation from days 5 to 9 (n = 46–52 blastocysts over 4 replicate studies). Panel c: IL6 supplementation from days 7 to 9 (n = 39–42 blastocysts/group over 3 replicate studies). Panel d: IL6 supplementation from days 5 to 7 then removal of embryos from IL6 treatments and culture in the absence of IL6 from days 7 to 9 (n = 29–42 embryos/group over 2 replicate studies). Different superscripts for each cell type within each panel indicate differences (P < 0.05)