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. 2015 Apr 20;25(4):391–400. doi: 10.1111/bpa.12161

Figure 1.

figure

Generation of p53‐null/Sparc‐wt and p53‐null/Sparc‐null mouse astrocytes. Homozygous p53‐null and Sparc‐null mice were mated to generate heterozygotes which were then mated to generate double‐null neonates. Four p53‐null/Sparc‐null astrocyte cell lines were generated and designated #2, #6, #11 and #30. Ast11.9‐2 is the control p53‐null/Sparc‐wt astrocyte cell line. A. PCR and B. Western blot analysis were used to confirm the loss of Sparc. The PCR product sizes are approximately 300 bp for the wt allele and 550 bp for the Sparc‐null insertion. The Western blot results show SPARC expression in lysate and conditioned medium (CM). C. Southern blot analysis was used to confirm the loss of p53. Knockouts were verified by loss of the wild‐type p53 band (∼5‐kb) and presence of the p53 mutant band (∼6.5‐kb).