Figure 1. Schematic diagram of the animal model used to generate intercaruncular endometria from heifers (n = 4), and lactating (n = 5), and nonlactating.

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Figure 1. Schematic diagram of the animal model used to generate intercaruncular endometria from heifers (n = 4), and lactating (n = 5), and nonlactating (n = 5) dairy cows on day 19 of pregnancy. FSH = follicle stimulating hormone; PG = prostaglandin analog; CIDR = controlled intravaginal drug-releasing device; AI = artificial insemination. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 2. Circulating concentrations of P4 (ng/ml) in heifers confirmed pregnant (black square), nonlactating cows confirmed pregnant (black circle), and lactating cows confirmed pregnant (black triangle) on day 19 following estrus. Serum P4 concentrations increased significantly in all three groups as pregnancy progressed (P< 0.05); however, there was no significant difference in P4 concentrations between lactating cows, nonlactating cows, and/or maiden heifers (P> 0.05). Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 3. Multidimensional scaling plot Figure 3. Multidimensional scaling plot. A multidimensional scaling plot (principal component analysis) for the top 500 genes showing the highest pairwise fold changes between the samples in the dataset was performed in EdgeR. L: lactating cows, NL: nonlactating cows, H: heifers. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 4. Hierarchical cluster analysis of DEGs found for lactating vs Figure 4. Hierarchical cluster analysis of DEGs found for lactating vs. nonlactating cows. Mean-centered expression values (log2 scale) were calculated for genes with P< 0.005 und used for hierarchical cluster analysis (MeV_4_8 v10.2). NL: nonlactating; L: lactating. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 5. Venn diagram for the overlap of DEGs between all three group comparisons, i.e. the differentially expressed genes that were common or distinct to each of the three comparisons made. L: lactating cows, NL: nonlactating cows, H: heifers. Venn diagram analysis was performed by jvenn [76] for the DEGs in each comparison that had a nominal P-value <0.001. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 6. Comparison of the differentially expressed genes in endometria of lactating vs. nonlactating cows with results from other studies. Genes with a P-value of < 0.005 were compared to lists of genes found as differentially expressed in bovine endometrium during the estrous cycle and early pregnancy as well as genes known or inferred to be regulated by estrogen. Red: increased expression, Blue: decreased expression (log2 fold), White: no match, L: lactating cows, NL, nonlactating cows, D: day. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 7. Mean glucose, lactate, and pyruvate concentrations in the uterine luminal fluid of lactating cows (black bar; n = 5 ± SD), dry cows (open bar; n = 6 ± SD), and maiden heifers (grey bar; n = 4 ± SD) confirmed pregnant on day 19. Effect of metabolic status on conceptus–maternal interactions on day 19 in dairy cattle: II. Effects on the endometrial transcriptome† Biol Reprod. Published online August 31, 2017. doi:10.1093/biolre/iox095 Biol Reprod | © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com