Figure 1.

Identification and phenotypic analysis of Samd4^PB/PB mice. (a) A schematic representation of the position of piggyBac (PB) transposon insertion into the Samd4 (sterile alpha motif domain containing protein 4) locus. The primers RT-F and RT-R were used for quantitative PCR (qPCR); and GL, GR and PBL were used for genotyping. Numbers 1–4 indicated exons 1–4. (b) Genotyping of littermates from two heterozygote cross by PCR. +/PB, the heterozygote mouse. (c) The reverse transcriptase-PCR analysis, which showed that Samd4 is widely expressed in the brain, heart, bone, white adipocyte and muscle, but little in the liver. (d) Analysis of Samd4 transcript levels by qPCR in wild-type mice (WT), Samd4^+/PB and Samd4^PB/PB tibia. (e) Western blotting analysis showing that Samd4 protein is absent in the protein from Samd4^PB/PB tibia. (f) Photograph of 3-week-old male WT and Samd4^PB/PB mouse. (g) Body weight of male WT and Samd4^PB/PB mice maintained from 1 to 12 weeks (n=21 for WT mice, n=17 for Samd4^PB/PB mice). (h) Kaplan–Meier survival curve (n=32 for WT mice, 24 for Samd4^PB/PB mice). Hprt, hypoxanthine guanine phosphoribosyl transferase; Tubulin, α Tubulin. Values represent means±s.d. P-values were obtained from t-tests with paired or unpaired samples, **P<0.01.