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Table 1.

Quantitative analysis of cellular Rh-EGF fluorescence

Measurement Nontransfected GFP-Rab21wt GFP-Rab21Q78L GFP-Rab21T33N
15 minutes      
   Structures/unit area   17.0±3.3   17.4±4.4   19.7±6.3   24.8±7.6** 
   Mean fluorescence/structure   140.3±17.1   139.8±19.8   127.1±18.9*  97.8±24.8*** 
   Total fluorescence of all structures (×103)   21.1±5.1   21.3±3.4   20.1±6.4   14.5±9.4* 
30 minutes      
   Structures/unit area   20.0±5.8   19.0±3.1   20.2±4.4   24.6±8.2  
   Mean fluorescence/structure   137.6±20.3   136.3±13.2   143.5±18.1   99.8±12.4*** 
   Total fluorescence of all structures (×103)   18.0±8.4   19.2±9.6   15.6±6.8   6.5±2.4*** 
Measurement Nontransfected GFP-Rab21wt GFP-Rab21Q78L GFP-Rab21T33N
15 minutes      
   Structures/unit area   17.0±3.3   17.4±4.4   19.7±6.3   24.8±7.6** 
   Mean fluorescence/structure   140.3±17.1   139.8±19.8   127.1±18.9*  97.8±24.8*** 
   Total fluorescence of all structures (×103)   21.1±5.1   21.3±3.4   20.1±6.4   14.5±9.4* 
30 minutes      
   Structures/unit area   20.0±5.8   19.0±3.1   20.2±4.4   24.6±8.2  
   Mean fluorescence/structure   137.6±20.3   136.3±13.2   143.5±18.1   99.8±12.4*** 
   Total fluorescence of all structures (×103)   18.0±8.4   19.2±9.6   15.6±6.8   6.5±2.4*** 

Quantitative analysis of Rh-EGF fluorescence in GFP-Rab21-expressing cells. HeLa cells transfected with GFP-Rab21 plasmids using calcium phosphate were incubated at 37°C for 15 and 30 minutes with 75 ng/ml Rh-EGF. The cells were washed on ice, fixed and analysed for the number of structures per unit area, the mean fluorescence per structure and the total fluorescence. Data shown are from ten cells, expressing moderate levels of either GFP-Rab21 wt, GFP-Rab21 Q78L, GFP-Rab21 T33N or nontransfected cells. At both time points, a similar number of Rh-EGF structures, with a similar fluorescence were observed in nontransfected cells and those expressing either GFP-Rab21 wt or GFP-Rab21 Q78L. However, cells expressing GFP-Rab21 T33N showed a distinct reduction in mean fluorescence per structure and in the total Rh-EGF fluorescence, despite containing more structures per unit area. Statistical significance is as follows

*

P<0.05

**

P<0.01

***

P<0.001 compared with nontransfected cells

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