Figure 3.
Simultaneous validation of multiple candidate genes involved in ATO/AsIII toxicity by secondary screening. A, Scatter plot showing relative enrichment (log2 FC > 0) or depletion (Log2 FC < 0) of each gene-specific guide sequence in ATO compared to vehicle control. Log2 fold changes (FC) are plotted against the average abundance of each guide sequence in the pool represented as Log2 counts per million (CPM). Hits with FDR values < 0.001 are shown in different colors. B, Functional enrichment analysis of validated genes showing the top biological processes affecting ATO toxicity. Bars represent numbers of validated genes within each pathway. C, Relative enrichment (Fold change > 1) of each of the guide sequences targeting KEAP1 in ATO-treated compared to control pools. D, Relative depletion (Fold Change < 1) of each of the guide sequences targeting ABCC1 in ATO-treated compared to control pools. Fold changes for each guide sequence are calculated by dividing the normalized abundance of the sequence in ATO-treated pools by that in control pools. The dotted line (y = 1) in each graph represents the baseline abundance of each guide sequence in control pools. Data are represented as mean (n = 3). Only guide sequences displaying differential abundance at FDR < 0.001 are shown.