Journal: Diabetologia

Article Title: Suppression of hyperinsulinaemia in growing female mice provides long-term protection against obesity

doi: 10.1007/s00125-015-3676-7

Figure Lengend Snippet: Attenuated obesity in HFD-fed Ins1 −/− : Ins2 +/− mice. ( a ) Body mass was tracked in pups ( n = 19–29, inset) and weaned mice ( n = 29–33, most time points). ( b ) DEXA-measured fat mass is shown ( n = 8–11). ( c , d ) Staining (magnification ×50) for perilipin (yellow) and DAPI (blue) in gonadal WAT of 25-week-old mice ( c ) is quantified ( n = 3) in ( d ) as frequency per size category. ( e , f ) Gene expression in gonadal ( e ) and inguinal ( f ) WAT mRNA is corrected against Hprt and normalised to CD-fed Ins1 −/− : Ins2 +/+ mice. Dark blue, CD-fed Ins1 −/− : Ins2 +/+ mice; dark purple, HFD-fed Ins1 −/− : Ins2 +/+ mice; light blue, CD-fed Ins1 −/− : Ins2 +/− mice; light purple, HFD-fed Ins1 −/− : Ins2 +/− mice. Data are means ± SEM. * p ≤ 0.05, ( * ) p = 0.051, CD vs HFD; † p ≤ 0.05, (†) p = 0.054, Ins1 −/− : Ins2 +/+ vs Ins1 −/− : Ins2 +/− ; * +/+ p ≤ 0.05, CD-fed vs HFD-fed Ins1 −/− : Ins2 +/+ mice; * +/− p ≤ 0.05, CD-fed vs HFD-fed Ins1 −/− : Ins2 +/− mice; †HFD p ≤ 0.05, HFD-fed Ins1 −/− : Ins2 +/+ vs Ins1 −/− : Ins2 +/− mice

Article Snippet: Insulin content was measured by ELISA from ten islets per mouse that were lysed at −20°C using 0.02 mol/l HCl in 70% ethanol and sonicated for 30 s. Other tissues were weighed and flash-frozen in liquid nitrogen before being stored at −80°C, except for half of the gonadal WAT depot, which was fixed in 4% paraformaldehyde, embedded in paraffin and cut into 5 μm-thick sections that were stained for perilipin (antibody 9349S; Cell Signaling Technology, Danvers, MA, USA) using an Alexa fluor 488-conjugated secondary antibody (Life Technologies, Burlington, ON, Canada) and DAPI.

Techniques: Staining, Gene Expression

Journal: Diabetologia

Article Title: Suppression of hyperinsulinaemia in growing female mice provides long-term protection against obesity

doi: 10.1007/s00125-015-3676-7

Figure Lengend Snippet: Long-term effects of HFD and INSULIN2. Mice were implanted with pumps releasing INSULIN2 or vehicle for 4 weeks (indicated by grey hatching). ( a ) Fasted insulin levels, n = 3. ( b ) Body mass, n = 3. ( c – e ) At 50 weeks, characterisation included WAT depot weight proportional to body mass ( c ) and gonadal WAT staining (magnification ×50) for perilipin (yellow) and DAPI (blue) ( d ) ( n = 3; quantification is shown in e as frequency per size category). ( f – h ) mRNA levels of genes expressed in gonadal WAT ( f ), inguinal WAT ( g ) and BAT ( h ) are corrected against Tbp or Hprt , and normalised to levels in Ins1 −/− : Ins2 +/+ mice. Dark purple, vehicle-treated Ins1 −/− : Ins2 +/+ mice; light purple, vehicle-treated Ins1 −/− : Ins2 +/− mice; hatched bars and triangles, INSULIN2-treated Ins1 −/− : Ins2 +/− mice. Data are means ± SEM. ‡ p ≤ 0.05, for the indicated comparisons; (‡) p = 0.058 in ( f ) for group effect; and (‡) p ≤ 0.057 in ( h ) for the indicated comparisons

Article Snippet: Insulin content was measured by ELISA from ten islets per mouse that were lysed at −20°C using 0.02 mol/l HCl in 70% ethanol and sonicated for 30 s. Other tissues were weighed and flash-frozen in liquid nitrogen before being stored at −80°C, except for half of the gonadal WAT depot, which was fixed in 4% paraformaldehyde, embedded in paraffin and cut into 5 μm-thick sections that were stained for perilipin (antibody 9349S; Cell Signaling Technology, Danvers, MA, USA) using an Alexa fluor 488-conjugated secondary antibody (Life Technologies, Burlington, ON, Canada) and DAPI.

Techniques: Staining