Fig. 4: The prepubertal injection of domestic cats with AAV9-fcMISv2 prevents breeding-induced ovulation and pregnancy, and lead to disrupted estrous behavior.

Female domestic cats were treated prepubertally with either 5 × 10¹² viral particles per kilogram of body weight of AAV9-Empty vector (Controls, black, circles), or 5 × 10¹² (Low-dose, blue, squares) or 1 × 10¹³ (High-dose, red, triangles) viral genomes per kilogram of body weight of AAV9-fcMISv2. a Inverted Kaplan-Meier survival plot showing the probability of pregnancy for each group during the mating trial. b Stacked histograms showing the average percentage of days that females from each group had at least one successful breeding attempt (black), no successful but at least one unsuccessful breeding attempt (gray) or no breeding interaction with a male (white). The percentage of days with successful breeding attempts was compared between the six treated females that bred and the two control females using an unpaired two-tailed T test with Welch’s correction: **P = 0.0097. c Average cumulative successful breeding attempts for each group of females. Data are shown as means ± SD. d Violin plots showing the distribution of the duration of breeding bouts observed during the mating trial. Dashed lines indicate the median, dotted lines indicate the quartiles. e Timeline of successful breeding activity, luteal phases, and pregnancies. Days with at least one successful breeding attempt are shown in black, blue, or red. Luteal phases are shown in gray. Days with either only unsuccessful breeding attempts or no breeding activity and without elevated fecal progestogen readings are shown in white. f Average fecal progestogen metabolite readings relative to the number of days since the first day of the preceding breeding bout. Progestogen metabolite levels were normalized to the first day of a breeding bout for each individual cat. A bell-shaped, non-linear regression curve was interpolated for the control group (black line). A simple linear regression curve was interpolated for the low- (blue line) and high- (red line) dose groups. n  =  2, 4, and 3 (a, e) and n = 2, 3, and 3 (b–d, f) female domestic cats in the control, low-dose, and high-dose groups, respectively. Source data are provided as a Source data file.