Fig. 3

Cdk4/6 inhibition by Palbociclib (PD) rescues the uhrf1 mutant liver phenotype. (A) Treatment scheme for inhibitor Palbociclib (PD). (B) Representative images of 120 hpf uhrf1−/− and control embryos after DMSO or PD treatment, liver indicated in red. (C) Quantification of the left liver lobe area of 120 hpf PD treated and untreated livers, segregated based on the liver size. Normal, medium and small liver size ranges are defined based on mean of DMSO treated controls and mutants ±1 standard deviation, as shown by the error bars. (D) Quantification of the number of hepatocytes at 120 hpf uhrf1−/− and control embryos after DMSO or PD treatment based on Z-stack images of larvae with the hepatocyte nuclear marker, Tg(fabp10a:nls-mCherry). (E) Quantification of 30 min and 12 min EdU incorporation at 120 hpf in DMSO and PD treated controls and uhrf1−/−. (F) Representative confocal image of 120 hpf Hoechst stained control and uhrf1−/− hepatocytes from DMSO or PD treatment and quantification of the nuclear surface area. (G) TUNEL assay was performed to detect cell death in 120 hpf in DMSO and PD treated controls and uhrf1−/− and quantified. (H) Genotype of all larvae that survived at 9 dpf following treatment with vehicle or PD from 48–120 hpf. The number of larvae that were treated and the number that remained at 9 dpf are indicated below the graph. (I) Representative slot blot for 5-mC and double stranded (ds) DNA in 120 hpf uhrf1−/− whole embryos and sibling controls, with quantification of the methylation levels from seven clutches. Scale: 1000 μm in B, 50 μm in F, the number of samples and clutches indicated for each condition and represented as median with range. P-value *< 0.05, **< 0.005, *** < 0.0005 by unpaired Student's t-test with adjustment for multiple comparisons.