Fig 1.
Deletion of atypical E2fs in mice has no effect on growth and survival of mice.
(A) Xgal staining of different organs derived from 9 weeks old CreERT2-/- E2f7LoxP/LoxP E2f8LoxP/LoxP R26R-LacZLoxP/LoxP mice (78R26Rf/f) and CreERT2+/- E2f7LoxP/LoxP E2f8LoxP/LoxP R26R-LacZLoxP/LoxP (78R26RΔ/Δ). Newborn pups of both groups were injected with tamoxifen for three days from postnatal day 2. (B) qPCR for E2f7 and E2f8 mRNA levels in indicated organs of 78f/f and 78∆/∆ mice aged 9 weeks. (C) Body weight measurements over time of 78f/f mice and 78∆/∆ mice. Bar graphs represents mean and standard error.
Fig 2.
E2f7/8 are essential for polyploidization in the pancreas.
(A) Hematoxylin & Eosin staining (400x magnification), top panel and DAPI (630x magnification, 1.5x zoom factor for exocrine, and 630x, 3x zoom factor for endocrine pancreas) lower panel, showing decreased nuclear sizes in 78∆/∆ compared to control 78f/f mice injected with tamoxifen at day 2 after birth and analysed at the age of 22 weeks. (B) Quantification of DAPI fluorescence intensity in pixels. (C) Representative flow cytometry histograms of pancreas tissues showing increased number of diploid cells and decreased number of tetraploid cells in 78∆/∆ compared to control 78f/f mice. (D) Summary histograms of flow cytometry profiles for each genetic group (n = 5). (E) Quantification of binucleation in exocrine pancreas; *p<0.05, bar graphs represents mean and standard error.
Fig 3.
Acute deletion of E2f7/8 in adult mice has no impact on polyploidy in the pancreas and liver.
(A) Beta catenin and DAPI staining of exocrine (630x magnification, 1.5 zoom factor) and endocrine pancreas (630x magnification, 3x zoom factor) and liver (400x magnification) showing similarity in nuclear sizes in the indicated genetic groups. Mice were injected with tamoxifen at the age of 8weeks and analyzed at the age of 22 weeks (B) Representative flow cytometry histograms of pancreas and liver showing similarity in diploid and tetraploid peaks. (C and D) Flow cytometry profiles in pancreas and liver showing similarity in percentage of diploid and tetraploid nuclei (n = 5). (E) qPCR for E2f7/8 in pancreas (n = 4); (F) qPCR for E2f7/8 in liver (n = 5). Bar graphs represents average and standard error; *p<0.05.
Fig 4.
Response to energy stress in E2f7/8-deficient pancreas.
(A) Immunostaining for insulin and glucagon under fed conditions in mice injected with tamoxifen at the age of 1 week and analysed at the age of 22 weeks. (B) Immunostaining of insulin and glucagon after 4hrs of starvation. (C-F) Serum biochemical parameters under normal feeding conditions. (G-J) Serum biochemical parameters following 4hrs of starvation. Bar graphs represents average and standard error, *p<0.05.
Fig 5.
Analysis of glucose, insulin, amylase and lipase in conventional E2f8 knockout mice under starving condition.
(A-D) Comparison of serum biochemical parameters after 12hrs of starvation. (E and F) Amylase protein in pancreas of indicated genotypes after 12hrs of starvation or 12hrs starvation. (G) Serum glucose levels after 2hrs of re-feeding. Bar graphs represents average and standard error, *p<0.05.