|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 4. Quantification of fibronectin endocytosis. (A) FN-null myofibroblasts were incubated with 2.5-50 µg/ml AF488-conjugated fibronectin at 4°C during the pulse phase (total endocytosis). Some samples were co-incubated with 800 µg/ml unlabeled fibronectin to determine non-specific endocytosis. Cells were chased in the absence of fibronectin for 2 hours at 37°C, and then processed for flow cytometry to quantify endocytosed fibronectin. Specific endocytosis was determined by subtracting nonspecific endocytosis from total endocytosis. The graph shows the best-fit curve of the mean fluorescence intensity (MFI) of internalized AF488-fibronectin. Data represent the mean of duplicate samples and error bars the range. (B) FN-null myofibroblasts were incubated with 10 µg/ml AF488-fibronectin in the presence of increasing concentrations of unlabeled fibronectin (0-800 µg/ml) during the pulse phase. Cells were chased in the absence of fibronectin for 2 hours at 37°C, and then processed for flow cytometry to quantify endocytosed AF488-fibronectin. The amount of internalized AF488-fibronectin is reported as percent control internalization (cells incubated in the absence of unlabelled fibronectin, which was set equal to 100%). Data represent the mean of duplicate samples and error bars indicate the range.
|
