1. first measured with iso-type control, that is, non-specific isotype antibody-stained cells, set a negative threshold.
2. Measure the cells transfected with the control shRNA using the specific antibody to see the percentage of positive cells
3. Measure the cells of the experimental group transfected with the shRNA to see the percentage of positive cells.
4. Finally, compare 2 and 3 to see if there is any change in the percentage of positive cells. If there is a significant decrease, the shRNA is effective. The percent change is the efficiency of the shRNA
5. If there is no significant change in the ratio of positive cells, but there is a shift in the fluorescence intensity of the cells, compare the median, or mean, of the fluorescence intensity of the positive cells