Fig. 5. Western blot analysis using cytoplasmic and nuclear extracts from U2Os cells transiently transfected with wild-type and mutant SHOX constructs. Immunofluorescence results using wild-type and mutant SHOX constructs were confirmed and complemented by western blots using cytoplasmic (Cy) and nuclear (Nu) extracts. Pyruvate kinase isozyme M2 (M2PK) at 57 kDa was used as a cytoplasmic marker and C23/nucleoloin at 116 kDa was used as a nuclear marker to detect the presence of SHOX (33 kDa) in both cytoplasmic or nuclear extracts.