(A). Representative Western blots (panel 1,3) and PCR analyzes (panel 2,4) of AIF expression in isolated mouse (panel 1,2) and human (panel 3,4) islets. Actin was used as loading control/ house keeping gene. Western blots/ PCRs are representatives of three independent experiments from three mice or from 3 organ donors, respectively. (B) Histological analysis by insulin staining in green and glucagon staining in red show a normal islet cellular composition and smaller islets in 2-week-old Hq mutant mice. (C,D) Analysis of β-cell mass (C) or β-cell mass divided by body weight (D) of WT andHq mutant mice at 2 and 9 weeks of age. Values are representative of 5 slides spanning the whole pancreas of each mouse and 4 mice for each group at each age (magnification x125). (E) Cell cycle characteristics of β-cells from WT mice and Hqmutant mice as measured by BrdU and pHH3 staining. BrdU⁺insulin⁺ cells are counted as β-cells at S phase (see example in F, upper panel). pHH3⁺ (with punctuated pattern) insulin+ cells are counted as β-cells at G2 phase (see example in F, Hq mice lower right panel). pHH3⁺ (with strong nuclear expression) insulin+ cells were counted as β-cells at M phase (see example in F, WT mice lower left panel). Data are shown as mean±SE. *P<0.05 in Hq mutant mice vs. WT mice.

https://doi.org/10.1371/journal.pone.0117766.g001