Figure 4
Loss of a single Cdk5rap3/Lzap allele in LZAP heterozygous mice results in increased resistance of cells to DNA damage. (a) LZAP protein levels in LZAP+/− and LZAP+/+ MEFs (genotyping of MEFs is shown in Supplementary Figure S2). (b) Caspase 3/7 activity (cleavage of the fluorescent substrate) in LZAP+/− and LZAP+/+ MEFs treated with zeocin for 6 h; experiment was performed twice in three LZAP+/− or three LZAP+/+ MEFs; P-values are calculated with unpaired t-test. (c) LZAP+/− and LZAP+/+ MEFs were treated with increasing concentration of carboplatin or zeocin, alive cells were visualized by methylene blue staining 7 days after the treatment. (d) Viability of MEFs from c was determined by methylene blue extraction, followed by quantification of absorbance. Percent survival is shown relative to control cells; error bars show s.e.; assays were performed in duplicate, P-values were calculated with paired t-test. (e) Survival of bone marrow progenitor cells derived from untreated wild-type or LZAP+/− mice or littermates treated with 6 Gy total body irradiation was determined after 7 days of in vitro growth. Data are presented as mean±s.d. (n=2 mice per group).
