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. 2005 Feb 3;24(4):717–729. doi: 10.1038/sj.emboj.7600559

Figure 1.

Figure 1

Identification and characterization of PACS-2, a sorting protein found on the ER and mitochondria. (A) Top: Schematic and Kyte–Doolittle hydrophobicity plot of the human PACS-1 and PACS-2 proteins. FBR, cargo/adaptor-binding region; ARR, atrophin-1-related region. Radiation hybrid and genome database analyses mapped the PACS-1 gene to chromosome 11q13.1 (Genbank AY320283) and the PACS-2 gene to chromosome 14q32.33 (Genbank AY320284). Bottom: Northern blot analysis of the tissue distribution for PACS-1 and PACS-2 transcripts. (B) Confocal immunofluorescence of endogenous PACS-1 and PACS-2 in A7 cells. PACS-1/PACS-2 were visualized with Alexa488 (green) and markers were visualized with Alexa546 or mitotracker (red). Scale bar, 10 μm. (C) A7 cells transfected with PACS-1 or PACS-2 siRNAs were analyzed by Western blot 48 h post-transfection. (D) A7 cells were transfected or not with PACS-1 or PACS-2 siRNAs. After 48 h, cells were processed for immunofluorescence microscopy using anti-CI-MPR (green) and anti-TGN46 (red). (E) A7 cells were transfected with control (scrambled), PACS-1, or PACS-2 siRNAs for 48 h and processed for confocal immunofluorescence localization of mitochondria (mitotracker, red) and ER (PDI, green). (F) A7 cells were transfected with the corresponding siRNAs and assayed for cell death by Annexin V/propidium iodide staining and FACS analysis. Treatment of cells with the proapoptotic CtBP siRNA served as a positive control (Zhang et al, 2003). Error for all graphs=s.d.