Research Paper Volume 16, Issue 20 pp 12977—13011

Figure 1. WRN associates with HDAC2. (A) Representative image of HDAC2/WRN PLA in the isogenic HDAC2+ and hdac2-27 null derivatives of GM639. Scale, 20 µm. (B) Quantitation of HDAC2/WRN PLA performed as in (A). (C) Hdac2-27 cells were stably transfected with HDAC-V5 transgene or empty vector (tag-less) and analyzed by Western blotting with antibodies to NCL (internal control) and V5. (D) Quantitation of IF in situ performed with V5 antibody on the same cells as in (C). MFI, mean fluorescence intensity, a.u., arbitrary units. (E) Representative image of V5/WRN PLA performed on the same cells as in (C, D) that were labeled for 30 min with EdU prior to harvest. EdU incorporation was visualized by Clicking to Alexa 488 azide. Scale, 20 µm. (F) Quantitation of V5/WRN PLA performed as in (E). V5/WRN signals of EdU-positive and EdU-negative cells are plotted separately. (G) WI38hTERT cells were stably transfected with the indicated shRNA-expressing constructs and treated with 100 ng/ml doxycycline for 5 days prior to Western blotting with WRN antibody. i.c., internal control (the STING protein). I.c.-normalized WRN levels are shown relative to shNS w/o DOX. (H) Quantitation of HDAC2/WRN PLA performed with the same cells as in (F). Crossbars in jitter plots are distribution means. Here and elsewhere, see Methods for details of p-value calculations.