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. 2013 Apr 23;4:1769. doi: 10.1038/ncomms2742

Figure 4. miR-155 impairs brown fat differentiation in vivo.

Figure 4

(a) qRT–PCR analysis for miR-155 expression in igWAT and BAT of 10-week-old male mice. Expression in igWAT was set as one; n=3. (b) Schematic representation of the lentiviral construct (LVPGK-miR-155) used for generation of miR-155 transgenic (miR155TG) animals. For abbreviations, see Supplementary Fig. S1, PGK, phosphoglycerate 1 promoter. (c) Interscapular BAT isolated from 1-week-old wt or miR155TG mice. Bright field (left) and fluorescent images (right) are shown; GFP is co-expressed with miR-155 in transgenic mice; (scale bar =2 mm). (d) Hematoxylin and eosin staining of BAT sections from 1-week-old wt and miR155TG littermates; (scale bar =15 μm). (e) Western blot analysis of UCP1, PGC-1α, PPARγ and C/EBPβ expression in BAT isolated from 1-week-old wt and miR155TG littermates; each protein levels from three representative animals per group are shown. Tubulin served as loading control. (f) Densitometric quantification of UCP1, PGC-1α, PPARγ and C/EBPβ expression levels, normalized to tubulin. Data of the wt group were set as one; n=6 per group. (g) Infrared thermographic analysis of the body surface temperature in 4-day-old wt and miR155TG littermates. Three representative pairs are showed. (h) Statistical analysis of body surface temperature in wt and miR155TG littermates; wt: 34.75±0.39 °C; miR155TG: 33.24±0.53 °C; n=5 per group. (i) Interscapular BAT, igWAT, and visceral WAT (vWAT) were isolated from 10-week-old wt or miR155TG littermates. Scale bar, 3 mm (BAT); 4 mm (igWAT and vWAT). Two representative littermates are shown. (j) Statistics of BAT, igWAT, vWAT, spleen, heart, brain and liver weights relative to total body weight in 10-week-old wt and miR155TG littermates; n=3 per group. (k) qRT–PCR analysis of brown thermogenic/adipogenic genes in interscapular BAT of wt or miR155TG littermates at 10 weeks of age: UCP1, PGC-1α and PPARγ; n=3 per group. All data are presented as means±s.e.m. (*P<0.05; **P<0.01; n.s., not significant; Student’s t-test).