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[Preprint]. 2026 Jul 23:2026.07.22.738304. [Version 1] doi: 10.64898/2026.07.22.738304

Figure 1: PCSK inhibition blocks follicular rupture in an ex vivo model of ovulation via effects on the COC.

Figure 1:

A) Schematic showing methodologies for testing the direct effect of PCI on antral follicles (top) and COCs (bottom). B) Left: Time-lapse images from videos of follicles cultured ex vivo with or without PCI. Scale bar = 100 μm. Right: Brightfield images of COCs removed from control or PCI-treated follicles. Scale bar = 25 μm. C) Has2 expression in follicles treated with PCI for 4 h. Data are represented as mean ± SEM. **** p < 0.0001 by two-way ANOVA. n=5-6 follicles per treatment group. D) Expansion velocity of CTL and PCI-treated COCs across in vitro maturation in the EmbryoScope+™. Data are represented as mean ± SEM. * p < 0.05 by mixed-effects analysis. n=48 biological replicates per treatment group. E) Representative brightfield images of COCs treated with PCI or DMSO control (CTL) following culture and transfer out of the EmbryoScope+™ dish. F) Representative brightfield images of residual cumulus cells in EmbryoScope+™ wells after removal of CTL or PCI-treated COCs. G) Graph showing COC area of CTL or PCI-treated COCs following transfer out of the EmbryoScope+™ dish. Data are represented as mean ± SEM. **** p < 0.0001 by Welch’s t-test. n=26 biological replicates per treatment group. H) Live/dead assay of COCs cultured in CTL or PCI-treated media for 16 h in the standard incubator. Live cells are shown in green (calcein AM) and dead cells are shown in red (ethidium homodimer). Scale bar = 200 μm. I) Quantification of live cumulus cells relative to total cumulus cells in CTL and PCI-treated COCs. Data are represented as mean ± SEM. p > 0.05 by Fisher’s exact test. n=2 biological replicates (5-14 pooled COCs per treatment group). J) Representative brightfield images of CTL vs. PCI treated COCs after 16 h culture and denuding. K) Graph showing maturation status of CTL or PCI-treated COCs after 16 h of culture. Data are represented as mean ± SEM. p>0.05 by Fisher’s exact test. n=15-16 biological replicates per treatment group. L) Immunofluorescent images depicting chromosomes (DAPI; blue), spindles (alpha-tubulin; green), and actin (rhodamine phalloidin; red) in CTL or PCI-treated COCs after denuding of cumulus cells. White asterisks denote spindles and yellow asterisks denote the polar body. Scale bar is 25 μm. Graphs showing percent of M) COCs with bipolar spindles and N) aligned chromosomes after culture with or without PCI. For M) and N), Data are represented as mean ± SEM, p>0.05 by Fisher’s exact test, and n=3 biological replicates (15-16 pooled COCs per treatment group).