Fig. 6.

SCAF1 is a new genome maintenance factor. A, the cell lines indicated were virally transduced with sgRNAs targeting SCAF1 and clonogenic survival was assessed in the presence or absence of olaparib (16 nM) on the pool of transduced cells. sgRNA targeting efficiency was assessed using genomic PCR amplification of the targeted locus and TIDE analysis. Editing efficiencies were >50% for all conditions (see Experimental Procedures). Data is represented as mean +SEM (n = 3), p-values are obtained using a two-tailed t test. B, same as (A), but here SCAF1 was depleted using siRNA transfection prior to clonogenic survival. Data is represented as mean +SEM (n = 3), p-value is obtained using a two-tailed t test. C, the cell lines indicated were transfected with the siRNAs indicated and after 24 h cells were exposed to IR (10 Gy). Cells were allowed to recover for 3 h, fixed and the proportion of RAD51 cells with greater than five foci was assessed by immunofluorescence. Data was acquired with a high-content screening station ScanR. Data from three independent experiments were combined; data are represented as mean ± SEM. D, Same as (C), expect that the experiment was performed by SN. Data is represented as mean +SEM (n = 3), p-value is obtained using a two-tailed t test. E, same as (C), except that the indicated cell lines were used. EV, empty vector; UT, untreated.