Figure 6: Loss of COC matrix integrity due to PCSK inhibition is associated with changes in cumulus cell migration behavior and adhesive properties.

A) Box plots of candidate genes related to migration with are upregulated with PCI treatment. 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. B) Representative brightfield images of CTL COCs stained with SPY-DNA 650 throughout IVM to track cumulus cell migration, with accompanying 3D projections. C) Representative displacement plots of CTL and PCI-treated COCs. Analysis was performed in Imaris (Bitplane) with tracks limited to those with duration greater than 6.5 h. D) Top: Mean square displacement plots of control and PCI-treated COCs, shown with ± SEM. Bottom: Graph showing the anomalous diffusion exponent ( ) of CTL vs. PCI-treated COCs relative to normal diffusion ( =1). E) Immunofluorescent images of actin (gray/red) and chromosomes (DAPI; blue) in control vs. PCI-treated COCs. Scale bar = 25 μm. F) Scanning electron microscopy images of CTL and PCI-treated cumulus cells. G) Violin plots of proteins related to cell adhesion 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, where each replicate has 150 COCs per treatment group.