Supplementary Figure 6: FAM35A and C20orf196 functions relating to homologous recombination.
From: Shieldin complex promotes DNA end-joining and counters homologous recombination in BRCA1-null cells
a, Representative images for quantifications of GFP-FAM35A presented in Fig 6a; scale bar 10µm. b, Representative images for quantifications presented in Fig 6b; scale bar 10µm. c, Representative images of FAM35A and C20orf196 effects on DNA-end resection in wild-type and BRCA1ko cells as measured by RPA nuclear intensity (after pre-extraction) following camptothecin treatment in the indicated genotypes, quantified in Supplementary Fig 6d; scale bar 10µm. d, Quantification of nuclear RPA intensity; n=5 independent experiments, except WT siAbraxas and BRCA1ko siCtIP (n=2) and BRCA1ko/FAM35Ako (n=4), with individual data points plotted over bars. e, Quantification of BRCA2 accrual at laser micro-irradiated RPE1 cells with the indicated genotypes for the representative images presented in Fig 6d. n=3 independent experiments, with individual data points plotted over bars. f, Gating strategy employed for TLR assay. g, AUC for clonogenic survival assay presented in Fig 6g. N=4 independent experiments, except BRCA1ko/FAM35Ako +FAM35A and +N-terminus where n=3, and +C-terminus where n=2; with individual data points plotted over bars. h, FAM35A and 53BP1 effects on olaparib resistance in BRCA1ko cells are not additive as measured by clonogenic survival assay (left panel), AUC (right panel). N=4 independent experiments, except BRCA1ko/53BP1ko/FAM35Ako where n=2; with individual data points plotted over bars. In d, e, g and h, bars represent mean ± SEM, one-way Anova; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns=not significant (p≥0.05); statistical source data including the precise p values can be found in Supplementary Table 5.
