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Fig. 3. Cellular expression of ALK-FH-derived proteins: normalization of the expression levels and subcellular localization. (A) Western blot analysis of the expression of ALK-FH-derived proteins after normalization of their expression levels. PC12 cells were electroporated with the indicated quantities of the DNA constructs, in order to obtain comparable levels of protein expression. Cells were lysed after two days in a RIPA buffer as described in the Materials and Methods, and the lysates (10 µg) were submitted to a western blot analysis using the anti-HA-tag (12CA5) monoclonal antibody. (B) Immunofluorescence confocal microscopy analysis of the subcellular localization of ALK-FH-derived proteins revealed with the anti-HA-tag antibody (green) in permeabilized PC12 cells (representative fields are shown). Cell nuclei were labeled with the Sytox Orange nucleic acid stain (red). Bar, 10 µm.
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