Supplementary Figure 3: Generation of IFFO1 and LMNA knockout DT40 cells.
a, Schematic representation of the chicken IFFO1 and targeted genomic DNA. The regions containing exon1-7 (marked by red) of the gene are replaced by blasticidine and histidinol resistant genes. The two regions between two pairs of dotted lines were used as arms of the knockout constructs. Arrows indicated targeted location of primers for genomic PCR. b, Genome-PCR analysis to show that IFFO1 gene is undetectable in the respective knockout DT40 cells. XLF was included as a positive control. c, Immunoblotting to show that IFFO1 protein is absent in the knockout cells. d, Sensitivity assay of the IFFO1-/- DT40 cells to various DSB-induced agents. Bars represent mean ± SD; n = 3 independent experiments. e, Schematic representation of the chicken LMNA and targeted genomic DNA. The regions containing exon2-7 (marked by red) of the gene are replaced by blasticidine, histidinol and puromycin resistant genes. The two regions between two pairs of dotted lines were used as arms of the knockout constructs. Arrows indicated targeted locations of primers for genomic PCR. f, Genome-PCR analysis to show that LMNA gene is undetectable in the respective knockout DT40 cells. XRCC4 was included as a positive control. g, Sensitivity assay of the LMNA-/-/- DT40 cells to various DSB-induced agents. Bars represent mean ± SD; n = 3 independent experiments. All immunoblots and gels are representative of three independent experiments; unprocessed scans of immunoblots are shown in Supplementary Fig. 9. Statistical source data including precise P values are shown in Supplementary Table 5.
