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Table 1 Phenotypes of transfected cell lines expressing inducible GFP-BLM alleles.

From: The C-terminal domain of the Bloom syndrome DNA helicase is essential for genomic stability

   

Localization

Relative

Relative

Cell Line

% green cells

% green

of GFP-BLM

Expression

Expression

 

no inducer

cells +Tet

fusion proteins

no inducer

+Tet

GFP

8

33

diffuse (NM1)

nd

nd

GFP-BLM

1

94

NBs + NM2

0.16(0.03)

1.0(0.15)

GFP-ΔN1

1

75

NM2 (3% NBs)

0.17(0.05)

0.96 (0.3)

GFP-ΔN2

9

90

NM2

0.30(0.08)

2.28 (0.35)

GFP-ΔN3

2

74

NM2 + BBs

0.05(0.02)

0.87(0.12)

GFP-ΔN4

1

77

NM1&2

0.09(0.01)

0.81 (0.09)

GFP-K695T

3

30

NBs + NM2 + BBs

0.12(0.01)

0.45 (0.08)

GFP-ΔH1

3

80

NBs +NM3

0.02(0.01)

0.39 (0.08)

GFP-ΔC1

1

77

NBs + BBs

0.04(0)

0.83 (0.03)

GFP-ΔC2

5

82

NBs + BBs

0.01(0)

0.74 (0.06)

  1. At least 200 DAPI-stained cells were counted and scored for green fluorescence to determine the percentages of induced and uninduced cells expressing the GFP-BLM fusion proteins. All GFP-fusion proteins show diffuse nuclear staining. The nucleolar morphologies (NM) of each fusion protein are reported as shown in Figure 4. NM1 is a diffuse overall nucleolar localization (Figure 4A &4H). NM2 is overall nucleolar fluorescence with exclusion from central holes " TOPIIIα. BBs are numerous spherical BLM-containing nuclear bodies that do not co-localize with either antigen. At least 300 cells were examined for determination of nuclear morphologies. Expression levels of the fusion proteins without Tet and with 1 μg/ml Tet relative to normal BLM are reported as determined in Figures 3 &5. The values reported are the average of two experiments. The standard deviation for these data points is shown in parentheses beside each value.