Fig. 4

Colocalization of BECN1 and BCL2L1. Oocytes that underwent either TN/TN IVM or HS/TN IVM were fixed and used for IHC to determine co-localization. Colocalization of BECN1 (green) and BCL2L1 (red) was low regardless of time point during IVM or temperature treatment. There appeared to be more punctate intense spots of BECN1 staining in oocytes having undergone 21 h of IVM under HS compared to the oocytes that underwent 21 h of IVM at TN conditions (A). The comparison between 21-h TN (B), 21-h HS (C), or negative control (D) shows that there was punctate green signal representing BECN1 but no colocalization in both TN and HS oocytes. For A, the white bar in images in the left and middle columns represent 50 μm, and the white bar in images in the right column represents 20 μm. The white arrows point to areas of punctate BECN1 signal but no colocalization. The images in the right column are magnified 5× images from the same oocyte from the middle column