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Table 1.

Cell-cycle-dependent enrichment of EBNA1 at oriP and the remote reference site I3

A. EBNA1
Cell cycle phases (Cp difference)
Average
Primer Econst. G1 G1/S S S/G2 G2 Cp Enrichment
sc2   1.74 ± 0.01   5.6 ± 0.14   4.8 ± 1.56   5.7 ± 0.78   5.3 ± 1.2   4.7 ± 1.0   5.2 ± 0.93   18.2 ± 4.3  
sc3   1.84 ± 0.01   8.5 ± 0.85   8.0 ± 1.06   9.0 ± 0.35   9.1 ± 1.34   8.1 ± 2.05   8.5 ± 1.13   185 ± 55  
sc4   1.83 ± 0.03   9.6 ± 0.07   9.9 ± 0.42   10.0 ± 1.13   9.9 ± 0.35   8.6 ± 1.1   9.6 ± 0.78   348 ± 82  
sc5   1.97 ± 0.03   9.7 ± 0.21   9.2 ± 1.48   9.9 ± 0.28   9.8 ± 0.99   9.2 ± 1.7   9.5 ± 0.93   659 ± 149  
sc6   1.95 ± 0.03   10.0 ± 0.42   8.4 ± 1.20   9.5 ± 0.14   9.4 ± 0.71   8.6 ± 1.27   9.1 ± 0.71   506 ± 199  
sc7   1.92 ± 0.03   9.0 ± 0.21   8.7 ± 1.20   9.3 ± 0.71   9.6 ± 0.49   8.7 ± 0.71   8.9 ± 0.71   371 ± 96  
sc8   1.94 ± 0.04   9.4 ± 0.57   8.4 ± 1.2   8.8 ± 1.06   8.9 ± 0.21   8.1 ± 1.56   8.8 ± 0.93   340 ± 90  
sc10   1.88 ± 0.02   3.4 ± 0.57   3.6 ± 0.38   3.4 ± 0.35   4.4 ± 0.42   4.3 ± 0.71   3.8 ± 0.47   11.5 ± 3.7  
I3   1.98 ± 0.01   3.0 ± 0.64   3.3 ± 0.49   2.8 ± 0.35   3.5 ± 0.35   3.3 ± 0.42   3.2 ± 0.28   8.8 ± 1.6  
A. EBNA1
Cell cycle phases (Cp difference)
Average
Primer Econst. G1 G1/S S S/G2 G2 Cp Enrichment
sc2   1.74 ± 0.01   5.6 ± 0.14   4.8 ± 1.56   5.7 ± 0.78   5.3 ± 1.2   4.7 ± 1.0   5.2 ± 0.93   18.2 ± 4.3  
sc3   1.84 ± 0.01   8.5 ± 0.85   8.0 ± 1.06   9.0 ± 0.35   9.1 ± 1.34   8.1 ± 2.05   8.5 ± 1.13   185 ± 55  
sc4   1.83 ± 0.03   9.6 ± 0.07   9.9 ± 0.42   10.0 ± 1.13   9.9 ± 0.35   8.6 ± 1.1   9.6 ± 0.78   348 ± 82  
sc5   1.97 ± 0.03   9.7 ± 0.21   9.2 ± 1.48   9.9 ± 0.28   9.8 ± 0.99   9.2 ± 1.7   9.5 ± 0.93   659 ± 149  
sc6   1.95 ± 0.03   10.0 ± 0.42   8.4 ± 1.20   9.5 ± 0.14   9.4 ± 0.71   8.6 ± 1.27   9.1 ± 0.71   506 ± 199  
sc7   1.92 ± 0.03   9.0 ± 0.21   8.7 ± 1.20   9.3 ± 0.71   9.6 ± 0.49   8.7 ± 0.71   8.9 ± 0.71   371 ± 96  
sc8   1.94 ± 0.04   9.4 ± 0.57   8.4 ± 1.2   8.8 ± 1.06   8.9 ± 0.21   8.1 ± 1.56   8.8 ± 0.93   340 ± 90  
sc10   1.88 ± 0.02   3.4 ± 0.57   3.6 ± 0.38   3.4 ± 0.35   4.4 ± 0.42   4.3 ± 0.71   3.8 ± 0.47   11.5 ± 3.7  
I3   1.98 ± 0.01   3.0 ± 0.64   3.3 ± 0.49   2.8 ± 0.35   3.5 ± 0.35   3.3 ± 0.42   3.2 ± 0.28   8.8 ± 1.6  
B.
x-fold enrichment above reference level (I3-fragment)
Primer Econst. G1 G1/S S S/G2 G2/M Average
sc2   1.74 ± 0.01   2.6   1.6   2.6   2.1   1.6   2.1 ± 0.5   
sc3   1.84 ± 0.01   24.6   17.4   32.7   34.8   21.7   26.2 ± 7.4   
sc4   1.83 ± 0.03   36.9   45.6   48.4   44.2   24.9   40.0 ± 9.5   
sc5   1.97 ± 0.03   79.8   56.9   94.6   88.4   58.8   75.7 ± 17.1   
sc6   1.95 ± 0.03   91.4   37.1   65.4   61.2   35.9   58.2 ± 22.9   
sc7   1.92 ± 0.03   39.4   32.4   49.6   58.3   33.5   42.7 ± 11.11   
sc8   1.94 ± 0.04   52.5   30.6   41.5   44.4   26.8   39.1 ± 10.4   
sc10   1.88 ± 0.02   1.0   1.1   1.0   1.8   1.7   1.3 ± 0.4   
B.
x-fold enrichment above reference level (I3-fragment)
Primer Econst. G1 G1/S S S/G2 G2/M Average
sc2   1.74 ± 0.01   2.6   1.6   2.6   2.1   1.6   2.1 ± 0.5   
sc3   1.84 ± 0.01   24.6   17.4   32.7   34.8   21.7   26.2 ± 7.4   
sc4   1.83 ± 0.03   36.9   45.6   48.4   44.2   24.9   40.0 ± 9.5   
sc5   1.97 ± 0.03   79.8   56.9   94.6   88.4   58.8   75.7 ± 17.1   
sc6   1.95 ± 0.03   91.4   37.1   65.4   61.2   35.9   58.2 ± 22.9   
sc7   1.92 ± 0.03   39.4   32.4   49.6   58.3   33.5   42.7 ± 11.11   
sc8   1.94 ± 0.04   52.5   30.6   41.5   44.4   26.8   39.1 ± 10.4   
sc10   1.88 ± 0.02   1.0   1.1   1.0   1.8   1.7   1.3 ± 0.4   

(A) Summary of the histogram in Fig. 2B. Econst was determined from standard curves of tenfold dilutions as described (Schepers et al., 2001). The Cp differences and standard deviations of three independent experiments are shown for the five fractions analysed. The mean values of each DNA segment and cell-cycle phase were determined (Cp difference). Because EBNA1 is cell-cycle-independently bound, the mean value of each fragment was established (Cp average) and used as exponent to determine the enrichment by the equation N=EconstΔCp (average enrichment). (B) To calculate the oriP-specific enrichment, we divided the mean value of the individual scanning fragments and the different cell cycle points with the accumulation of the reference fragment I3 (ΔCp=3.2 cycles, Econst=1.98, enrichment=1.983.2=8.8 times). The average enrichments over the cell cycle are listed on the right.

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