Additional file 1 of Obesity alters the mouse endometrial transcriptome in a cell context-dependent manner

Additional file 1: Supplemental Figure 1. Increased macrophages in the high-fat diet endometrium. Supplemental Figure 2. Purification of endometrial cells. Supplemental Figure 3. Differential gene expression analysis of high-fat diet uterine macrophages. Supplemental Figure 4. Impact of estrous cycle stage on differential gene expression. Supplemental Figure 5. Negative immunohistochemistry staining

Fluorine-Substituted Pyrrolo[2,3‑<i>d</i>]Pyrimidine Analogues with Tumor Targeting via Cellular Uptake by Folate Receptor α and the Proton-Coupled Folate Transporter and Inhibition of <i>de Novo</i> Purine Nucleotide Biosynthesis

Novel fluorinated 2-amino-4-oxo-6-substituted pyrrolo­[2,3-<i>d</i>]­pyrimidine analogues <b>7</b>–<b>12</b> were synthesized and tested for selective cellular uptake by folate receptors (FRs) α and β or the proton-coupled folate transporter (PCFT) and for antitumor efficacy. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> showed increased <i>in vitro</i> antiproliferative activities (∼11-fold) over the nonfluorinated analogues <b>2</b>, <b>3</b>, <b>5</b>, and <b>6</b> toward engineered Chinese hamster ovary and HeLa cells expressing FRs or PCFT. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> also inhibited proliferation of IGROV1 and A2780 epithelial ovarian cancer cells; in IGROV1 cells with knockdown of FRα, <b>9</b>, <b>11</b>, and <b>12</b> showed sustained inhibition associated with uptake by PCFT. All compounds inhibited glycinamide ribonucleotide formyltransferase, a key enzyme in the <i>de novo</i> purine biosynthesis pathway. Molecular modeling studies validated <i>in vitro</i> cell-based results. NMR evidence supports the presence of an intramolecular fluorine–hydrogen bond. Potent <i>in vivo</i> efficacy of <b>11</b> was established with IGROV1 xenografts in severe compromised immunodeficient mice

Fluorine-Substituted Pyrrolo[2,3‑<i>d</i>]Pyrimidine Analogues with Tumor Targeting via Cellular Uptake by Folate Receptor α and the Proton-Coupled Folate Transporter and Inhibition of <i>de Novo</i> Purine Nucleotide Biosynthesis

Novel fluorinated 2-amino-4-oxo-6-substituted pyrrolo­[2,3-<i>d</i>]­pyrimidine analogues <b>7</b>–<b>12</b> were synthesized and tested for selective cellular uptake by folate receptors (FRs) α and β or the proton-coupled folate transporter (PCFT) and for antitumor efficacy. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> showed increased <i>in vitro</i> antiproliferative activities (∼11-fold) over the nonfluorinated analogues <b>2</b>, <b>3</b>, <b>5</b>, and <b>6</b> toward engineered Chinese hamster ovary and HeLa cells expressing FRs or PCFT. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> also inhibited proliferation of IGROV1 and A2780 epithelial ovarian cancer cells; in IGROV1 cells with knockdown of FRα, <b>9</b>, <b>11</b>, and <b>12</b> showed sustained inhibition associated with uptake by PCFT. All compounds inhibited glycinamide ribonucleotide formyltransferase, a key enzyme in the <i>de novo</i> purine biosynthesis pathway. Molecular modeling studies validated <i>in vitro</i> cell-based results. NMR evidence supports the presence of an intramolecular fluorine–hydrogen bond. Potent <i>in vivo</i> efficacy of <b>11</b> was established with IGROV1 xenografts in severe compromised immunodeficient mice

Fluorine-Substituted Pyrrolo[2,3‑<i>d</i>]Pyrimidine Analogues with Tumor Targeting via Cellular Uptake by Folate Receptor α and the Proton-Coupled Folate Transporter and Inhibition of <i>de Novo</i> Purine Nucleotide Biosynthesis

Novel fluorinated 2-amino-4-oxo-6-substituted pyrrolo­[2,3-<i>d</i>]­pyrimidine analogues <b>7</b>–<b>12</b> were synthesized and tested for selective cellular uptake by folate receptors (FRs) α and β or the proton-coupled folate transporter (PCFT) and for antitumor efficacy. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> showed increased <i>in vitro</i> antiproliferative activities (∼11-fold) over the nonfluorinated analogues <b>2</b>, <b>3</b>, <b>5</b>, and <b>6</b> toward engineered Chinese hamster ovary and HeLa cells expressing FRs or PCFT. Compounds <b>8</b>, <b>9</b>, <b>11</b>, and <b>12</b> also inhibited proliferation of IGROV1 and A2780 epithelial ovarian cancer cells; in IGROV1 cells with knockdown of FRα, <b>9</b>, <b>11</b>, and <b>12</b> showed sustained inhibition associated with uptake by PCFT. All compounds inhibited glycinamide ribonucleotide formyltransferase, a key enzyme in the <i>de novo</i> purine biosynthesis pathway. Molecular modeling studies validated <i>in vitro</i> cell-based results. NMR evidence supports the presence of an intramolecular fluorine–hydrogen bond. Potent <i>in vivo</i> efficacy of <b>11</b> was established with IGROV1 xenografts in severe compromised immunodeficient mice

Tumor Targeting with Novel Pyridyl 6‑Substituted Pyrrolo[2,3‑<i>d</i>]Pyrimidine Antifolates via Cellular Uptake by Folate Receptor α and the Proton-Coupled Folate Transporter and Inhibition of <i>De Novo</i> Purine Nucleotide Biosynthesis

Tumor-targeted specificities of 6-substituted pyrrolo­[2,3-<i>d</i>]­pyrimidine analogues of <b>1</b>, where the phenyl side-chain is replaced by 3′,6′ (<b>5</b>, <b>8</b>), 2′,5′ (<b>6</b>, <b>9</b>), and 2′,6′ (<b>7</b>, <b>10</b>) pyridyls, were analyzed. Proliferation inhibition of isogenic Chinese hamster ovary (CHO) cells expressing folate receptors (FRs) α and β were in rank order, <b>6</b> > <b>9</b> > <b>5</b> > <b>7</b> > <b>8</b>, with <b>10</b> showing no activity, and <b>6</b> > <b>9</b> > <b>5</b> > <b>8</b>, with <b>10</b> and <b>7</b> being inactive, respectively. Antiproliferative effects toward FRα- and FRβ-expressing cells were reflected in competitive binding with [³H]­folic acid. Only compound <b>6</b> was active against proton-coupled folate receptor (PCFT)-expressing CHO cells (∼4-fold more potent than <b>1</b>) and inhibited [³H]­methotrexate uptake by PCFT. In KB and IGROV1 tumor cells, <b>6</b> showed <1 nM IC₅₀, ∼2–3-fold more potent than <b>1</b>. Compound <b>6</b> inhibited glycinamide ribonucleotide formyltransferase in <i>de novo</i> purine biosynthesis and showed potent <i>in vivo</i> efficacy toward subcutaneous IGROV1 tumor xenografts in SCID mice

Maximum Likelihood tree based on the 3 concatenated mtDNA genes (COI, COII and cyt <i>b</i>) (1527bp).

<p>Values above branches correspond to MP and ML bootstrap values (only values > 50% are shown) and values below branches correspond to Bayesian posterior probability. PT – Portugal; SP – Spain; UK – United Kingdom; BL – Belgium; FN – Finland; SC – Sicily; TK – European Turkey; AT – Anatolia; USA – United States of America; CA – Canada.</p

Median-joining haplotype network of <i>Philaenus spumarius</i> sampled geographic regions for mitochondrial gene COI (539bp).

<p>Size of the circles is in proportion to the number of haplotypes. Branches begin in the centre of the circles and their sizes are in proportion to the number of mutations.</p

Maximum Parsimony tree based on nuclear gene elongation factor-1α.

<p>Values above branches correspond to MP bootstrap (only values > 50% are shown) and Bayesian posterior probability values. Black: GenBank sequences (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098375#pone.0098375-MaryanskaNadachowska1" target="_blank">[26]</a>); blue individuals correspond to the eastern mtDNA group; red individuals correspond to the eastern-Mediterranean mtDNA group and green individuals to the western-Mediterranean mtDNA group.</p

Sampling locations of <i>Philaenus spumarius</i> in (a) Europe and Anatolia (b) North America and (c) New Zealand in each geographic region.

<p>1 – Azores; 2 – Iberian Peninsula; 3 – Morocco; 4 – France; 5 – United Kingdom; 6 – Belgium; 7 – Italian Peninsula; 8 – Sicily; 9 – Slovenia; 10 – Balkans (Bulgaria; Greece; European Turkey); 11 – Anatolian Peninsula; 12 – Finland; 13 – North America (Canada and United States of America); 14 – New Zealand. Circle sizes are proportional to the number of individuals. Circles: green – “western-Mediterranean” mtDNA group; red – “eastern-Mediterranean” mtDNA group; blue – “eastern” mtDNA group. Circle sizes are proportional to the number of samples.</p

Tajima's <i>D</i> and [44] Fu's <i>F</i>s test values and their statistical significance for <i>Philaenus spumarius</i> Cytochrome <i>c</i> oxidase I mtDNA groups.

<p>*: indicates significant values at P<0.05; **: indicates significant values at P<0.01 and ***: indicates significant values at P<0.001.</p