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. 2019 Apr 6;16:407–418. doi: 10.1016/j.omtn.2019.03.008

Figure 6.

Figure 6

Effect of hAEC-Derived Exosomes on Chemotherapy-Induced Granulosa Cell Apoptosis In Vitro

(A) hAECs were labeled with PKH26 fluorescent dye. Schematic of the experimental procedure used to harvest PKH26-labeled exosomes from the PKH26-hAEC-conditioned medium. PKH26-labeled hAEC exosomes were added to the media of KGN cells. (B) The fluorescence signals were detected by confocal microscopy in KGN cells. The nuclei of the hAECs and KGN cells were stained with DAPI (blue) and their cytoskeleton with Actin-Tracker Green (green). Scale bar, 40 (A) and 10 (B) μm. (C) Western blot analyzed the proliferation marker PCNA and the apoptosis-related proteins expression in KGN cells in the different treatment groups. (D) qRT-PCR was used to detect miR-1246, miR-197-5p, miR-1290, miR-4516, miR-4281, and miR-21-5p expression in granulosa cells. (E) The level of miR-1246 and miR-21-5p expression was detected in granulosa cells transfected with miR-1246 and miR-21-5p mimic and inhibitors. (F) Western blot was performed to detect PCNA and cleaved Caspase 3 protein expression in granulosa cells. *p < 0.05 versus Control or NC group; #p < 0.05 versus the CTX and inhibitor NC group.