Figure 1
Deficiency in tuberin is associated with significant decrease in NF-YA and loss of OGG1 expression in kidney tumor of Eker rats. A. Immunoblot analysis of tuberin, NF-YA and OGG1 protein expression in normal kidney of wild type rats and tumor kidney tissue from Eker rats. Actin was used as loading control. B. Histograms represent means ± SE (n = 3). Significant difference from wild type rat is indicated by ** P < 0.01.
Figure 2
Downregulation of tuberin expression in human renal epithelial cells results in decrease in NF-YA and OGG1 expression. A. Immunoblot analysis of tuberin, NF-YA and OGG1 in HEK 293 cells transfected with siRNA directed against TSC2 for 48 h. Actin was used as a loading control. B. Histograms represent means ± SE (n = 2). Significant differences from cells transfected with the TSC2-specific siRNA are indicated by **P < 0.01.
Figure 3
Mislocalization of NF-YA in tuberin-deficient cells. A. TSC2-/- and TSC2+/+ cells were plated on a 2-well chamber slide. Cells were fixed then immunostained with primary anti-NF-YA antibody and secondary anti-goat IgG labeled with Alexa Fluor 488 (red fluorescence). The same cells were also incubated with DAPI (blue fluorescence, same microscopic fields) to identify nuclei. Overlay of NF-YA and DNA staining, demonstrating nuclear localization of NF-YA in the wild type and tuberin-deficient cells infected with adenovirus expressing tuberin. B. Nuclear and cytoplasmic fraction proteins were extracted from Tsc2-/- cells non-infected and infected with adenovirus expressing tuberin. Immunoblot analysis of tuberin and NF-YA in the nuclear and cytoplasmic fractions was analyzed as described in Method section. Lamin B and actin antibodies were used as nuclear and cytoplasmic markers, respectively.