Figure S2.
Characterization of UOK262 Cells Complemented with FH and FHcyt, Related to Figure 4
(A-C) Complementation with, and localization of, FH and FHcyt in UOK262 cells (Yang et al., 2010b) was confirmed by immunofluorescence. The FH and FHcyt constructs carry a C-terminal V5 tag (green); mitochondria are identified with Mitotracker red (red) and nuclei by Dapi (blue).
(A) Parental UOK262 cells show no V5 immunofluorescence. Four panels are shown; three for each fluorophore separately and a merged image.
(B) UOK262 cells transfected with FH show clear overlap with Mitotracker red confirming the mitochondrial localization.
(C) Cytoplasmic localization of FHcyt was confirmed since there is no overlap with Mitotracker red. The large vacuoles that are present in these cells can also be seen clearly in this set of images.
(D) Complementation of UOK262 cells with FH and FHcyt was confirmed further by Western blot analysis for FH and the C-terminal V5 tag. ACTB was used as a loading control. Note that complementation with either transgene did not ameliorate HIF-1α levels.