FIG. 1.

Identification and downregulation of all PAT proteins (ADFP and Tip47) in AML12 cells. A: Cells were incubated with 400 μmol/l oleic acid for 12 h before staining. Cells were costained with a polyclonal goat anti-ADFP antibody and with a rabbit polyclonal anti-Tip47 antibody and, respectively, alexa fluor 488–or 594–conjugated secondary antibodies. Fluorescent and phase images were generated by an LSM 510 confocal laser microscope. Bar represents 50 μm. B: Coimmunostaining with ADFP and Tip47 of AML12 cells treated for 4 days with siRNA ADFP, siRNA Tip47, both combined, or control. Cells were treated as above. C: Immunoblots of total cellular protein extract from AML12 cells treated with control siRNA (Qiagen) (lane 1); siRNA ADFP (lane 2); siRNA Tip47 (lane 3); or both combined (lane 4). Rabbit polyclonal anti-Tip47, anti-ADFP, and anti–β-actin antibodies were used as loading control. (Please see http://dx.doi.org/10.2337/db07-1383 for a high-quality digital representation of this figure.)