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  1. sFig. 1 sFig. 1Telomeric repeat amplification protocol (TRAP) showing that gemcitabine does not affect telomerase activity. Hela cells were treated with gemcitabine(1.0 mg/ml) for 3 days and then subjected to TRAP assays with cell numbers of 100, 250 and 500 for individual drug concentration. 500* indicates the samples of cell number 500 that were treated with heat at 95o C for 5 min. A buffer control was added to show no contamination in our reagent. IC represents internal control. 0 1.0 GMT (g/ml) Buffer 500* 500* 100 250 500 100 250 500 - IC

  2. sFig. 2A shTRF2 stable lines V #1 #2 #3 #4 #5 + – + + + + + – – – – – - TRF2 - Actin 21226 5148 4973 4268 3530 2027 1904 1584 Loading control

  3. sFig. 2B sFig. 2. TRF2 is required for maintaining telomere integrity. Helacells were transfectedwith shTRF2 plasmid and selected with puromycin. Resistant clones were expanded for mass culture. The TRF2 levels were tested by Western blot with anti-TRF2 antibody (A, upper panel). Five TRF2-repressed clones and a vector control were treated with gemcitabine1 mg/ml for 3 days and then subjected to TRF length assay (A, lower panel) . Panel B shows relative TRF2 level (B, left) and TRF length (B, right) with or without gemcitabinetreatment . 2.5 Mean TRF length (kbp) Relative TRF2 expression GMT (–) GMT (+) GMT (–) GMT (+)