Figure 5: Molecular changes due to PCSK inhibition translate into phenotypic changes in COC matrix organization.

A) Schematic showing composition of COC ECM, with components of interest in bold. B) Box and whisker plots showing expression of key COC ECM genes including Has2, Vcan, and Ptx3 in CTL vs. PCI treated COCs. Data are represented as mean ± SD. Adjusted p-values for box plots as follows: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001 by Wilcoxon signed-rank test. n=3 biological replicates of 10 pooled COCs per treatment and timepoint. C) Violin plots of proteins related to matrix remodeling that are altered with PCI treatment. Data are represented as mean ± SEM. q-values for violin plots as follows: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001 by t-test with Storey’s method. n=4 biological replicates of 150 pooled COCs per treatment and timepoint. D) Representative brightfield images of CTL or PCI-treated COCs cultured for 12 in agarose micromolds and stained with hematoxylin and eosin. Boxes highlight locations where high magnification images were captured. Arrows point to edge of ECM in CTL and PCI-treated COCs. E) Graph showing comparison of cumulus cell count between CTL vs. PCI-treated COCs. Data are represented as mean ± SEM. p > 0.05 by Welch’s t-test. n=10-18 COCs across 4 trials. F) Immunofluorescent images of hyaluronan stained with hyaluronic acid binding protein (HABP; green) and chromosomes (DAPI; blue) in control vs. PCI-treated COCs. Scale bar = 25 μm. **** p < 0.0001 by Welch’s t-test. G) Box plot of Cemip expression in CTL vs. PCI-treated COCs across in vitro maturation. * p < 0.05 by Wilcoxon signed-rank test. n=3 biological replicates of 10 pooled COCs per treatment and timepoint.