Figure 3.

Assessing the accuracy of RBMB-based measurements of RNA copy number. HT1080-GFP-96mer cells were microporated in the presence of RBMBs and placed in the incubator for 5 h. The concentration of RBMBs was varied from 0.05 μM to 4.8 μM. The cells were then fixed and smFISH was performed. (A) A custom Matlab program was used to determine the percent of RBMB-reporter signals that were colocalized with smFISH signals and the number of smFISH signals that were colocalized with RBMB-reporter signals, on a cell-by-cell basis. Each data point represents the mean measurement of colocalization (±standard deviation) for at least 20 cells, collected from at least two independent experiments. (B) Measurements of RNA copy number per cell, as determined by smFISH and RBMBs, were also compared.