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. 2019 Dec 19;8:e47430. doi: 10.7554/eLife.47430

Figure 1. Genome-wide screen identifies TLE3 as a modulator of AR inhibitor sensitivity.

(A) Overview of the genome-wide CRISPR-Cas9 resistance screen. (B) Representation of the relative abundance of the gRNA barcode sequences of the CRISPR-Cas9 resistance screen. The y-axis shows the enrichment (relative abundance of apalutamide treated/untreated) and the x-axis shows the average sequence reads of the untreated samples. (C) Long-term growth assay (14 days) showing the functional phenotype of LNCaP cells harboring TLE3 knockout or knockdown vectors, cultured in the presence of vehicle or enzalutamide. Cells harboring a non-targeting sgRNA (sgNT) or scrambled shRNA (shSCR) were used as a control. (D) Quantitative analysis of live cell proliferation in real-time for control cells and TLE3KO cells in the absence or presence of enzalutamide. (E) Western blot showing TLE3 protein levels for control cells and TLE3KO cells shown in C and D. Vinculin was used as a loading control.

Figure 1—source data 1. Normalized readcounts CRISPR screen.
Normalized readcounts obtained with massively parallel sequencing are shown for each condition of the CRISPR screen (with three replicates per condition): timepoint 0, untreated and ARN-treated cells.
Figure 1—source data 2. DESeq2 analysis of the CRISPR screen.
Output file showing the raw data obtained with DESeq2 for the CRISPR resistance screen. For each gRNA the results, as indicated at the top of each column, are shown.
Figure 1—source data 3. MAGeCK analysis of the CRISPR screen.
Output file showing the raw data obtained with MAGeCK for the CRISPR resistance screen. For each gRNA the results, as indicated at the top of each column, are shown.

Figure 1.

Figure 1—figure supplement 1. Genome-wide CRISPR screen identifies TLE3 as a modulator of AR inhibitor sensitivity.

Figure 1—figure supplement 1.

(A–B) Long-term growth assay of LNCaP cells treated with apalutamide or enzalutamide for 14 days. Enzalutamide-resistant 22rv1 cells were used as control. (C) MAGeCK analysis showing hits obtained from the CRISPR-Cas9 resistance screen. (D) TLE3 mRNA expression levels in LNCaP cells harboring shRNAs targeting TLE3. Cells with scrambled shRNA (shSCR) were used as a control. (E) Long-term growth assays showing the functional phenotype for control and TLE3KO LNCaP cells cultured in the presence of vehicle or apalutamide at indicated concentrations for 14 days. (F–G) Long-term growth assays showing the functional phenotypes for indicated cell lines carrying control or TLE3-targeting gRNAs , cultured in the presence of vehicle or enzalutamide at indicated concentrations. (H) Western blot analysis showing TLE3 expression levels for the cells shown in F and G. GAPDH was used as a loading control.