Number, size and aspect of lipid droplets are affected by overexpression of miR-30b during lactation.

<p>A/ Lipid droplets staining with Nile Red (green) and nuclei staining with DAPI (blue) on frozen histological sections of mammary gland from transgenic (Tg) and wild-type (WT) mice at day-12 of lactation. B/ The relative number of lipid droplets to number of nuclei, counted using ImageJ, is significantly lower in transgenic (Tg) than in wild-type (WT) mice. C/ The total area (measured in pixels with ImageJ) of the lipid droplets relative to their number is significantly higher in transgenic (Tg) than in wild-type (WT) mice. Data represent means ± S.E. obtained from day-12 lactating mammary glands frozen sections of 8 transgenic and 6 wild-type mice (10 pictures analyzed per individual). a, b: indicate a significant difference among lines (p<0.05, ANOVA).</p

miR-30b overexpression affects the mammary gland structure during lactation and involution.

<p>Histological analyses (HES) of mammary gland from transgenic (Tg) and wild-type (WT) mice at day-12 of lactation (A) and day-6 of involution (B).</p

miR-30 family targets validated in the literature.

<p>miR-30 family targets validated in the literature.</p

Apoptosis and proliferation processes are modified by overexpression of miR-30b during lactation.

<p>A/ Apoptosis is evaluated by TUNEL analysis: the number of TUNEL-positive cells (red) relative to the nuclei (blue) is significantly higher in transgenic (Tg) than in wild-type (WT) mice. B/ Cell proliferation activity is detected by Ki67 immunostaining: Ki67 labeled cells (green) are detected in transgenic mammary gland (Tg) samples whereas it is absent in wild-type (WT) ones. Nuclei (blue) were stained with DAPI and myoepithelial cells (red) were stained with α-SMA. Data represent means ± S.E. obtained from day-12 lactating mammary glands paraffin sections (5 pictures analyzed per individual) of 3 transgenic and 3 wild-type mice for TUNEL analysis and 6 transgenic and 7 wild-type mice for Ki67 immunostaining. a, b: indicate a significant difference among lines (p<0.05, ANOVA).</p

Genes showing the greatest degree of change at involution day-6.

<p>Genes showing the greatest degree of change at involution day-6.</p

Ingenuity Pathway Analysis-Top Networks during lactation day-12.

<p>Ingenuity Pathway Analysis-Top Networks during lactation day-12.</p

Ingenuity Pathway Analysis-Top Networks-Involution day-6.

<p>Ingenuity Pathway Analysis-Top Networks-Involution day-6.</p

miR-30b expression changes during mouse mammary gland development.

<p>Relative expression of miR-30b was determined by RT-qPCR at different physiological stages. miR-30b expression was normalized to <i>U6</i> expression. Virgin 4 weeks (V04), 8 weeks (V08), Gestation (G), Lactation (L) and Involution (I) time points (in days). Values are means ± S.E. (n = 3 technical repetitions on pool of 3 mice).</p

The most down and up regulated genes during lactation day-12.

<p>The most down and up regulated genes during lactation day-12.</p

Growth of the pups is affected by overexpression of miR-30b.

<p>The weights of pups fed by dams of Tg12 line (solid line) were compared to pups fed by control wild-type mice (dotted line) from day-1 through to day-16 after birth. Here the average of weight per pup from 3 or more litters of 8 animals per dam is presented. From day-3 (*) the difference between the two curves is significantly different (p<0.05). At day-15 this difference averages 37%.</p