Fig. 5

Individual target validation by flow cytometry. Flow cytometry analysis of control and candidate pgRNAs at T0, T3 and T6 after induction of transdifferentiation. A Flow cytometry plots of intergenic negative control, two positive controls targeting ratCEBPa and SPI1, and two protein coding targets FURIN and NFE2. CD19 B-cell marker is represented on the X-axis and Mac1 macrophage marker is represented on the Y-axis. Cells that do not undergo transdifferentiation remain in the Q4 quadrant (positive for CD19 -X-axis- and negative for Mac1 -Y-axis) (the percentages of cells in this quadrant are shown). B Percentage of cells with delayed transdifferentiation (Q4 quadrant) observed in controls and individually validated candidates for two biological replicates (R1 and R2) at T3 and T6 after induction of transdifferentiation. For lncRNAs LINC02432 and MIR3945HG we only have data for one biological replicate at T6. Average (Avg) and standard deviation (SD) between replicates, and two-tailed p-values (comparing the delayed population from each individual target and the intergenic negative control) are also shown. Most of the selected candidates show significant delay compared to the intergenic negative control at T3. Although the value observed for FURIN is not statistically significant, the magnitude of the delay indicates that it is a strong candidate to perform further validations