Fig. 3. Transfection of COS7 cells and keratinocytes with periplakin C-terminal constructs. (A-L) COS7 cells; (M-P) primary human epidermal keratinocytes. After transfection, the cells were fixed without prior extraction (A,E-G,I,M-P) or extracted with 0.6% saponin for 30 minutes before fixation (B-D,H,J-L). B and C are the same field; J-L are the same field; M and N are the same field; and O and P are the same field. See Fig. 1 for details of each construct. Green fluorescence: anti-HA (detects tagged periplakin constructs). Red fluorescence: anti-vimentin in COS7 cells; anti-keratin (LP34) in keratinocytes. Purple/blue fluorescence: nuclei stained with TOTO-3, Bars, 10 µm (A-C,E,F,I); 50 µm (D,G,H); 5 µm (J-P).