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. 2010 Jun 25;18(1):72–79. doi: 10.1038/cdd.2010.77

Figure 1.

Figure 1

PML levels parallel those of SIRT1. (a) Extracts from SIRT1−/−, WT, or SIRT1-tg MEFs were prepared and analyzed by western blotting using anti-PML antibodies. (b) Heart tissue derived from SIRT1−/−, WT, or SIRT1-tg mice was subjected to immunohistochemistry staining using anti-PML (Sigma, HPA008312) or anti-SIRT1 (Sigma, S5196) antibodies. (c) HeLa cells were transfected with siRNA against SIRT1. At the indicated times after transfection, cells extracts were analyzed by western blotting using anti-PML or anti-SIRT1 antibodies. (d) HeLa cells were transfected with siRNA against SIRT1 and at 48 h were subjected to immunostaining using anti-PML or anti-SIRT1 antibodies followed by confocal microscopy. PML staining in a representative image of cells expressing different SIRT1 levels is shown. Arrows indicate cells in which SIRT1 expression is diminished. (e) HEK-293 cells were transfected with the indicated expression plasmids, and 48 h after transfection cells extracts were analyzed by western blotting using anti-PML or anti-SIRT1 antibodies. Cell extract from untransfected cells (NT) was used as a control. (f) SIRT1−/− MEFs were transfected with the indicated expression plasmids, and 48 h after transfection cells extracts were analyzed by western blotting using anti-PML or anti-SIRT1 antibodies. Cell extract from untransfected cells (NT) was used as a control. The numbers given under the panel show densitometric data after normalization to actin, and compared with control (set as 1)