Figure 3
Increased apoptosis correlated with induction of pro-apoptotic 19ARF mRNA and of p53 target genes in Mysm1-deficient BM and thymi. (a) qPCR was performed with primers specific for p19ARF or p16Ink4a using cDNA from total thymocytes of Mysm1−/− (KO) or controls (wild type (WT)). Significantly increased p19ARF mRNA level in Mysm1−/− compared with WT thymocytes relative to GAPDH mRNA detected by qPCR. Results represent mean fold changes of three independent experiments, with two thymi of each genotype. (b) Gel electrophoresis of representative Mysm1−/− and WT PCR samples. (c) qPCR detection of p53 target genes p21Waf/Cip, Bax, Puma, and Noxa in LSK cells from Mysm1−/− BM and (d) in total Mysm1−/− thymocytes compared with controls relative to GAPDH. (e) Decreased relative Mcl-1 mRNA expression in Mysm1−/− BM. (f) qPCR performed with primers specific for indicated genes on Mysm1−/− (KO) or control (WT) cDNA from total thymocytes with GAPDH as the reference gene. Bar graphs show mean expression values ±S.D. of at least three independent experiments
