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. 2023 Jan 25;299(3):102949. doi: 10.1016/j.jbc.2023.102949

Figure 9.

Figure 9

Cellular localization of HsUMPS. HeLa cells, transfected with plasmid expressing either flag-tagged native HsUMPS protein (WT) or the flag-tagged oligomerization deficient mutant (S87TQ90E), were fractionated using the REAP approach (see Experimental procedures) to determine the localization of HsUMPS. For both WT and the mutant, HsUMPS appears predominantly in the cytoplasm but also localizes to the nucleus. GAPDH is used for a cytoplasmic marker while Nup98 is used for a nuclear marker. A and B, the experiments were conducted in either nucleotide rich growth media (A) or in nucleotide depleted media (B). C, the mutant proteins (C) show a similar cellular distribution. For all three sets of data, the first three (A and B) or four (C) lanes are for the whole cell fraction, the next three (A and B) or four (C) are for cytoplasmic fraction, and the last three (A and B) or four (C) are for the nuclear fraction. HsUMPS, human UMP synthase; REAP, rapid, efficient, and practical; UMP, uridine 5′-monophosphate.