Primer Sequences and Qiagen QuantiTect Primer Assays™.

<p><b>Abbreviations:</b> HPRT, Hypoxanthine-guanine phosphoribosyltransferase; BMAL1, brain and muscle arnt-like protein 1; Clock, circadian locomotor output cycles kaput; Cry1 and Cry2, cryptochrome 1 & 2; XBP1s/ XBP1t, x-box binding protein spliced & total; GRP78, 78 kDa glucose-regulated protein or Binding immunoglobulin protein(BiP); SREBP1c, Sterol Regulatory Element-Binding Protein 1c; PER1 and PER2, period 1 & 2 genes; SIRT1, sirtuin 1; <i>NFκB (RelA)</i>, nuclear factor kappa-light-chain-enhancer of activated B cells; IL-1β, interleukin 1-beta; NLRP3, NLR family, pyrin domain containing 3, NFKBIA, nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha; GLUT5, facilitated glucose/fructose transporter member 5.</p><p>Primer Sequences and Qiagen QuantiTect Primer Assays™.</p

Prenatal fructose exposure increases hepatic lipid content, and modifies transcription factors and fatty acid oxidative enzymes in neonates in a sex dependent manner.

<p>Offspring liver triglyceride levels and SIRT1, SREBP1c, PPARa, mRNA levels at 2 timepoints. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. Two-Way ANOVA Main Effects are indicated in text where the 2 factors are maternal diet (fructose) and offspring sex. Tukey’s <i>post-hoc</i> analyses are indicated by letters, where bars with different letters indicate significance p < 0.05. Control offspring are in open bars, fructose exposed offspring are in black bars. E21: embryonic day 21, P10: postnatal day 10. n = 6 per group per sex.</p

Offspring blood biochemistry.

<p>Data are presented as mean ± SEM. Values with different letters indicates significant differences. NS: not significant; inter’n: interaction; NEFA: non-esterified fatty acid; ALP: Alkaline phosphatase; AST: Aspartate aminotransferase; ALT: alanine aminotransferase, protein: total protein.</p><p>Offspring blood biochemistry.</p

Prenatal fructose exposure increases fructose transport and decreases fructose metabolism in offspring.

<p>Offspring GLUT5, fructokinase, PEPCK mRNA and liver glycogen levels at 2 timepoints. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. Two-Way ANOVA Main Effects are indicated in text where the 2 factors are maternal diet (fructose) and offspring sex. Tukey’s <i>post-hoc</i> analyses are indicated by letters, where bars with different letters indicate significance p < 0.05. Control offspring are in open bars, fructose exposed offspring are in black bars. E21: embryonic day 21, P10: postnatal day 10. n = 6 per group per sex.</p

Prenatal fructose intake significantly alters gene expression of maternal hepatic fructose and glucose metabolic enzymes.

<p>Maternal GLUT5, fructokinase, PEPCK mRNA and liver glycogen levels at 2 timepoints. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. * p < 0.05 compared to control fed mothers; n = 10 CON, n = 9 FR. CON: Control fed mothers, FR: Fructose fed mothers. E21: embryonic day 21, P10: postnatal day 10. GLUT5: fructose transporter, PEPCK: phosphoenolpyruvate carboxykinase.</p

Free fatty acid metabolic genes regulated by prenatal fructose exposure in female neonatal livers.

<p>Data are presented as fold change from control values. (-) indicates a decrease in gene expression.</p><p>Free fatty acid metabolic genes regulated by prenatal fructose exposure in female neonatal livers.</p

Maternal hepatic NLRP3 inflammasome and inflammatory signalling.

<p>Offspring NLRP3 inflammasome and IL-1β mRNA levels at 2 timepoints. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. Two-Way ANOVA Main Effects are indicated in text where the 2 factors are maternal diet (fructose) and offspring sex. Tukey’s <i>post-hoc</i> analyses are indicated by letters, where bars with different letters indicate significance p < 0.05. Control offspring are in open bars, fructose exposed offspring are in black bars. E21: embryonic day 21, P10: postnatal day 10. n = 6 per group per sex. NLRP3: NOD-like receptor family, pyrin domain containing 3, IL1β: interleukin 1 beta.</p

Prenatal fructose exposure induces hepatic ER stress in neonates at postnatal day 10.

<p>Offspring GRP78 and XBP1 mRNA levels at 2 timepoints. XBP1 is expressed as a ratio of spliced to total mRNA levels. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. Two-Way ANOVA Main Effects are indicated in text where the 2 factors are maternal diet (fructose) and offspring sex. Control offspring are in open bars, fructose exposed offspring are in black bars. E21: embryonic day 21, P10: postnatal day 10. n = 6 per group per sex.</p

Prenatal fructose intake induces maternal hepatic endoplasmic reticulum (ER) stress.

<p>Maternal GRP78 and XBP1 mRNA levels at 2 timepoints. XBP1 is expressed as a ratio of spliced to total mRNA levels. Data are presented as means ± S.E.M. All mRNA levels are relative to the geometric mean of housekeeping genes. * p < 0.05 compared to control fed mothers; n = 10 CON, n = 9 FR. CON: Control fed mothers, FR: Fructose fed mothers. E21: embryonic day 21, P10: postnatal day 10. GRP78: 78 kDa glucose-regulated protein, XBP1: X-box binding protein 1.</p

Maternal blood biochemistry.

<p>Data are presented as mean ± SEM.</p><p>* p<0.05 compared to control. NEFA (uM): non-esterified fatty acid; ALP: Alkaline phosphatase; AST: Aspartate aminotransferase; ALT: alanine aminotransferase.</p><p>Maternal blood biochemistry.</p