Skip to main content
. 2023 Sep 6;621(7978):415–422. doi: 10.1038/s41586-023-06506-6

Extended Data Fig. 9. related to Fig. 4. Polθ forms nuclear bodies (NBs) in G1.

Extended Data Fig. 9

a, b, Representative images of Polθ nuclear bodies (Polθ NBs) and 53BP1 nuclear bodies (53BP1 NBs) in cyclin A-negative (G1) cells. c, Size and intensity of Polθ foci in indicated cell cycle phases. From left to right, n = 98, 15, 86, 41. d, Trajectories of individual Polθ NBs in BRCA2−/− RPE-1 cells blocked in G1 (Palbociclib treatment). Representative images of Polθ NBs from formation at mitotic exit (flattening) to dissolution in G1 (PCNA-negative cells) in BRCA2−/− cells. e, Representative images and quantification of 53BP1 NBs in WT and POLQ−/− cells upon indicated treatment. From left to right, n = 45, 46, 53, 47. f, Representative images and quantification of 53BP1 NBs in BRCA2−/− cells following Polθ depletion (IAA treatment). From left to right, n = 43, 29. g, h, Representative images and quantification of Polθ NBs in FANCD2-negative (G1) cells upon indicated treatment. i, Immunoblot analysis at indicated time points upon Polθ degradation (IAA treatment). In (g), from left to right, n = 561, 448 (4, 2 replicates). j, Quantification of micronuclei (MN) positive for γH2AX and/or CREST (centromere marker) in BRCA2−/− cells following Polθ degradation (IAA treatment). From left to right, n = 75, 57; among two replicates. k, Quantification of Polθ foci in micronuclei (MN) in indicated cell lines. Scale bars represent 5 μm, except where indicated. Data represents three biological replicates, except where indicated. Data shows mean +/− S.E.M., except box plots (c) showing median with quartiles. For (c,f,j) two-tailed Mann-Whitney test; for (e) unpaired t-test was performed.