Journal: Nature Communications

Article Title: The CIP2A-TOPBP1 axis facilitates mitotic DNA repair via MiDAS and MMEJ

doi: 10.1038/s41467-025-65594-2

Figure Lengend Snippet: a Representative images and dot plot showing the number of TOPBP1–MUS81 colocalising foci in RPE1 p53 -/- FRT/TR WT or CIP2A -/- prometaphase cells following 400 nM aphidicolin for 18 h and synchronisation with 60 ng/ml nocodazole for 2 h (WT: n = 78; CIP2A -/- Cl.1: n = 79; Cl.2: n = 80; three independent experiments). b Representative images and dot plot of EdU foci in prometaphase RPE1 p53 -/- WT or CIP2A -/- cells after following 400 nM aphidicolin followed by 30 min synchronisation with 60 ng/ml nocodazole (WT: n = 77; Cl.1: n = 69; Cl.2: n = 74; three experiments). c EdU foci in prometaphase DLD1 BRCA2 -/- cells treated with siCTRL or siCIP2A (siCTRL: n = 75; siCIP2A: n = 74; three experiments; significance determined by two-tailed unpaired t-test). d Bar plot showing break-induced replication (BIR) efficiency after sgRNP-CAS9 cleavage of an I-SceI site in the pBIR-GFP reporter U2OS cell line treated with siCTRL, siPOLD3, siCIP2A, or siSLX4 (four experiments). e Representative images and dot plot of CIP2A–γH2AX colocalising foci in RPE1 p53 -/- FRT/TR WT cells untreated or treated with 400 nM aphidicolin for 18 h followed by synchronisation with 60 ng/ml nocodazole for 2 h (UT: n = 82; Aph: n = 69; three experiments; two-tailed unpaired t-test). f Representative images and dot plot of FANCD2–CIP2A colocalising foci in RPE1 p53 -/- WT cells under the same conditions as ( e ) (UT: n = 75; Aph: n = 70; three experiments; two-tailed unpaired t-test). g eGFP-Polθ foci in prometaphase DLD1 BRCA2 -/- eGFP-Polθ cells treated with doxycycline (100 ng/ml, 24 h) 48 h after siRNA treatment with siCTRL or siCIP2A for (siCTRL: n = 86; siCIP2A: n = 83; three experiments; two-tailed unpaired t-test). h Bar plot of alternative end-joining (Alt-EJ)/microhomology-mediated end joining (MMEJ) efficiency after sgRNP-Cas9 cleavage of an I-SceI site in EJ2 reporter U2OS cells treated with siCTRL, siPolθ, or siCIP2A (three experiments). In ( a , b , c , e , f , g ), grey dots represent individual measurements, black dots indicate medians per experiment, and bars show mean ± SEM. Scale bars: 10 µm. Statistical significance: a, b, d, and h by one-way ANOVA; others by two-tailed unpaired t-test. Source data are provided as a file.

Article Snippet: U2OS cells (RRID: CVCL_0042) stably integrated with EJ2 (RRID:Addgene_44025 ), a gift from J. M. Stark and pBIR (RRID:Addgene_49807 ), a gift from T. D. Halazonetis, were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% foetal bovine serum (FBS).

Techniques: Two Tailed Test