FIG 4.

Flow cytometry example plots of the HIV RNA Flow-FISH assay and the correlation between measured and expected frequencies of infected cells in spiking experiments. (A) The p24⁺ gag mRNA⁺ ACH-2 cells before and after induction with PMA/ionomycin. (B) Highest frequency of reactivated ACH-2 spiked into the CEMx174 line. (C) False positive event rate in pure CEMx174 cells. (D) Theoretical expected frequency of 1.5 to 1,500 events per million cells and experimental frequencies of HIV⁺ events experimentally identified by HIV mRNA/protein co-staining.