Additional file 2: of Lack of cortistatin or somatostatin differentially influences DMBA-induced mammary gland tumorigenesis in mice in an obesity-dependent mode

Malignancy features (proliferation rate, inflammation and de novo angiogenesis) analysis of 7, 12 dimethylbenz[α]anthracene (DMBA)-induced mammary gland tumors in low fat (LF)-fed and high fat (HF)-fed wild-type (WT), cortistatin (CORT)-knockout (KO) and somatostatin (SST)-KO mice. Values represent mean ± standard error of the mean of percentage of KI67-positive cells (a), number of mitotic cells (per each 10 high power fields (HPF)) (b), scored inflammatory status evaluating number of inflammatory cells (C) and scored de novo angiogenesis evaluating number of small blood vessels in a subset of the developed tumors (d). Inflammation and de novo angiogenesis were valued as 1/3 (scarce), 2/3 (moderate) or 3/3 (abundant). n.d. not determined due to the lack of available tumors or the tumoral piece to be evaluated. (PDF 33 kb

Specific set of primers for amplification of mouse transcripts used for quantitative real-time RT-PCR.

<p><i>Bp</i> base pairs; <i>Sn</i> sense; <i>As</i> antisense.</p><p>Specific set of primers for amplification of mouse transcripts used for quantitative real-time RT-PCR.</p

Additional file 1: of Lack of cortistatin or somatostatin differentially influences DMBA-induced mammary gland tumorigenesis in mice in an obesity-dependent mode

Histopathological analysis of 7, 12 dimethylbenz[α]anthracene (DMBA)-induced mammary gland (MG) tumors. Representative images and histopathological classification of DMBA-induced MG tumors formed in low fat (LF)-fed and high-fat (HF)-fed wild-type (WT), cortistatin (CORT)-knockout (KO) and somatostatin (SST)-KO mice. a Adenocarcinoma. b Well-differentiated squamous carcinoma. c Moderately differentiated squamous carcinoma. d undifferentiated carcinoma. Scale (top right) indicates 100 μm. (PDF 277 kb

Impact of diet-induced obesity on the expression of GH-IGF-I axis in the mammary fat pads of CORT-KO, SST-KO and their respective control mice.

<p>mRNA levels of GH-R, IGF-I, IGF-II, IGF-IR and PRL-R were measured by qPCR. Values represent means ± SEM of the mRNA copy number of each transcript adjusted by the normalization factor (n = 5–6). Global differences by 2-way ANOVA are shown at the top of each graphic (G: genotype effect; D: diet effect I: effect of interaction between genotype and diet; *, p<0.05; **, p<0.01; ***, p<0.001). Asterisks above the bars (*, p<0.05; **, p<0.01; ***, p<0.001) indicate significant differences between groups by Bonferroni post-hoc.</p

Impact of diet-induced obesity on the expression of SST/CORT/receptor subtypes in the mammary fat pads of CORT-KO, SST-KO and their respective control mice.

<p>mRNA levels of sst1, sst2, sst3, sst4 were measured by qPCR. Values represent means ± SEM of the mRNA copy number of each transcript adjusted by the normalization factor (n = 5–6). Global differences by 2-way ANOVA are shown at the top of each graphic (G: genotype effect; D: diet effect I: effect of interaction between genotype and diet; *, p<0.05; **, p<0.01; ***, p<0.001). Asterisks above the bars (*, p<0.05; **, p<0.01) indicate significant differences between groups by Bonferroni post-hoc.</p

Plasma leptin levels in WT, CORT-KO and SST-KO mice fed under LF and HF diets.

<p>Values represent mean ± SEM of ng/ml of plasma leptin. Global differences by 2-way ANOVA are shown at the top of each graphic (G: genotype effect; D: diet effect I: effect of interaction between genotype and diet; *, p<0.05; **, p<0.01; ***, p<0.001). Asterisks above the bars (*, p<0.05) indicate significant differences between groups by Bonferroni post-hoc.</p

Impact of diet-induced obesity on the expression of ghrelin system in the mammary fat pads of CORT-KO, SST-KO and their respective control mice.

<p>mRNA levels of ghrelin, In2-ghrelin, GOAT were measured by qPCR. Values represent means ± SEM of the mRNA copy number of each transcript adjusted by the normalization factor (n = 5–6). Global differences by 2-way ANOVA are shown at the top of each graphic (G: genotype effect; D: diet effect I: effect of interaction between genotype and diet; *, p<0.05).</p

Characterization of GH/IGF-I axis and the regulatory SST/CORT/ghrelin/receptors system in mammary fat pads of female mice (expressed as absolute copy number/50 ng total cDNA).

<p>Values represent means ± SEM of the mRNA copy number of each transcript (n = 5–6 mice).</p