| | | 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) |
- 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.