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. Author manuscript; available in PMC: 2010 Jul 10.
Published in final edited form as: Cell. 2009 Jul 10;138(1):90–103. doi: 10.1016/j.cell.2009.06.021

Figure 1. Conditional deletion of mouse TRF1.

Figure 1

(A) Schematic of the mouse TRF1 locus on chromosome 1 (NCBI locus ID 21749), the targeting construct, and the altered alleles of TRF1. R: EcoRI, P: PvuII, H: HindIII. F1. F2 and R: PCR primers.

(B) TRF1 PCR on tail DNA from mice of the indicated genotypes using the F1 and F2 forward primers and the R reverse primer. PCR products: wild type, 100 bp; flox, 152 bp; null allele, 500 bp.

(C) Immunoblot monitoring loss of TRF1 upon Cre-treatment of TRF1F/F MEFs. TRF1 (Ab1449) was detected 3 days after Cre treatment of TRF1F/F and TRF1F/+ MEFs. -Cre: mock infection, γ-tubulin: loading control.

(D) IF-FISH to monitor TRF1 at telomeres of TRF1F/F MEFs at day 3 post Cre. TRF1 IF, red; telomeric FITC PNA probe, green.

(E) Graph representing proliferation of TRF1-deficient MEFs.

(F) Phase-contrast microscopic images of primary TRF1F/F MEFs before and after Cre treatment. Cells were stained for SA-β-galactosidase at day 7 post Cre.

(G) Metaphase spreads showing the fragile telomere phenotype in TRF1 null cells. Telomeres highlighted by FISH (green) and DNA was stained with DAPI (red) at day 4 post Cre.