Fig. 1.
Developmental exposure to doxycycline exerts lasting effects on offspring body weight and gut morphology. a Experimental timeline to assess the lasting effects of developmental exposure to doxycycline on offspring outcomes. b C57Bl/6J male offspring body was changed significantly over time (two-way ANOVA, main effect of time, F6, 175 = 557.3, P < 0.0001.) and between treatment groups (two-way ANOVA, main effect of treatment, F1, 175 = 11.15, P= 0.001). c C57Bl/6J female offspring body weight was significantly changed over time (two-way ANOVA, main effect of time, F6,132= 606.0, P<0.0001), but not between treatment groups (two-way ANOVA, main effect of treatment, F1,22= 0.0002955, P=0.9864). d Body weight experiments with animals of a mixed C57BL/6J:129S1/SvImJ background, male offspring body weight was significantly changed over time (two-way ANOVA, main effect of time, F6, 186= 1191, P < 0.0001) and across treatment (two-way ANOVA, main effect of treatment, F1, 31= 14.57, P = 0.0006). e In the mixed background experiment, female offspring body weight was significantly changed across time (two-way ANOVA, mean effect of time, F 1, 127 = 204.5, P < 0.0001) and across treatment (two-way ANOVA, main effect of treatment, F6,127= 10.50, P =0.0017). f Following cessation of doxycycline exposure, the cecal weights of mixed background males and females were greater compared with control males and females at postnatal day 28 (t6= 2.844, P = 0.0294). g This effect was resolved by postnatal day 63 Male (t17 = 1.758, P = 0.0967) Female (t9 = 0.1532, P = 0.8816). Data represented as mean ± SD. * P < 0.05, ** P < 0.01, ***P < 0.001. C57 body weight experiment: Control male n = 9; Dox male n = 18; Control female n = 14; Dox female n = 10. Mixed background body weight experiment: Control Male n= 13; control female n= 8; Dox male n = 20; Dox female n = 11. Cecal Weight experiments: postnatal day 28 n = 4; postnatal day 70-90, male n = 9-10; female = 5