Extended Data Fig. 7: Shieldin is recruited to DSBs later than DYNLL1 and in G1 phase only. Related to Fig. 6.

(a) Representative images from Fig. 6a. (b) Quantification of GFP positive stripes for cells expressing DYNLL1-EGFP or SHLD1-EGFP after exposure to laser microirradiation and fixed at indicated time points. (c) Quantification of number of DYNLL1 foci for RPE1 cells transduced with lentivirus comprised of the Fucci system reporter assay. Cells were exposed to 10 Gy irradiation, fixed 6 h later, and processed using antibodies against Geminin, Cdt1, and DYNLL1. (d) Representative images for Fig. 6c and Extended Data Fig. 6c. (e) Representative western blots showing knockdown of indicated proteins for Fig. 6d. (f) RPE1 BRCA1^−/− were subjected to 10 Gy IR and 4 h later were fixed and processed for immunofluorescence using antibodies against SHLD1, Cyclin A, and γH2AX. (a-f) n = 3 biologically independent experiments, counting ≥ 100 cells per experiment. Error bars represent the mean±s.e.m. P-values for foci quantification and ‘laser positive’ cell analysis was calculated using two-sided unpaired t-tests. P values are indicated by nonsignificant (P >0.05), *(P < 0.05), **(P < 0.01), ***(P < 0.001), ****(P<0.0001). Black line in dot plots represent median. Scale bar = 20μm.