Ag11395

The SV40-transformed WRN-deficient fibroblast cell line (AG11395) was obtained from Coriell Cell Repositories (Camden, NJ, USA). To produce stable cell lines, AG11395 (WS) fibroblasts were transduced with retroviruses expressing the full-length cDNA encoding wild-type WRN (WS^WT), exonuclease-dead (WS^E84A) or helicase-dead (WS^K577M) (21 (link)). All the cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM; Life Technologies) supplemented with 10% FBS (Boehringer Mannheim) and incubated at 37°C in a humidified 5% CO₂ atmosphere.

The SV40-transformed WS cell line (AG11395) was obtained from Coriell Cell Repositories (Camden, NJ, USA). The AG11395 cells carries an Arg368 stop mutation that results in a truncated protein that is degraded and undetectable. To produce stable cell lines, AG11395 (WS) fibroblasts were transduced with lentiviruses expressing the full-length cDNA encoding wild-type WRN (WRN^WT), S1133A-WRN (WRN^S1133A) or S1133D-WRN (WRN^S1133D). The WRNWT and the AG11395 complemented with a helicase-dead WRN (WRN^K577M) have been described elsewhere54 (link). HEK293T cells were from American Type Culture Collection. HEK293TshWRN, cells were generated after transfection with pRS-puro-shWRN (5′-AGGCAGGTGTAG- GAATTGAAGGAGATCAG-3′; sequence ID: TI333414 Origene) and puromicin selection. All the cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM; Life Technologies) supplemented with 10% FBS (Boehringer Mannheim) and incubated at 37 °C in a humidified 5% CO₂ atmosphere. All cell lines have been tested for the presence of mycoplasma by PCR and DAPI-IF.