Figure 4
From: SIRT6 facilitates directional telomere movement upon oxidative damage
SNF2H is recruited to damaged telomeres and partially recovers damage-induced telomere movements in SIRT6 KO cells. (a) Telomeres of MEF SIRT6 KO cells transiently expressing GFP-Vector/GFP-SNF2H and KR-TRF1 for telomere tracking/damage induction were traced directly after induction using a 559 nm laser light full-power scan; n = 100 telomeres, p < 0.001. (b) U2OS cells overexpressing GFP-SNF2H and DsRed-TRF1 (DR-TRF1) or KR-TRF1 were imaged 36 hr after transfection and directly after damage induction, and images were merged. (c) Rate of co-localization of GFP-SIRT6 with DR-TRF1 or KR-TRF1 after light illumination for 20 min was calculated; n = 100, p < 0.001. (d) Rate of co-localization of GFP-SNF2H with KR-TRF1 after 20 min light illumination at damaged telomeres in MEF WT and SIRT6 KO cells; n = 100, p < 0.001. (e) FLAG-SNF2H, GFP-H133Y SIRT6, and KR-TRF1 were expressed in SIRT6 KO cells and illuminated with light for 20 min. H133Y SIRT6 is localized at KR-TRF1 as marked and rescues the recruitment of SNF2H in SIRT6 KO cells. (f) GFP or GFP-H133Y SIRT6 was transfected in KR-TRF1 stably expressed 293 cells and illuminated with light for 20 min before immnoprecipitation. After pull down by GFP-antibody, WB of SNF2H and GFP are shown. SNF2H was pulled down by GFP-H133Y SIRT6.
